断裂内含肽及其用途

技术领域

本发明属于生物技术领域，具体涉及断裂内含肽及其用途。

背景技术

众所周知，不能封装在用于基因治疗的AAV(腺相关病毒)中的基因可以断裂成两个片段。因此，每个片段都应该融合到断裂内含肽中，并在它们各自的AAV中传递。感染目标细胞后，内含肽将使得能够产生所需的目标蛋白(参见图1)。

然而，该方法的主要限制之一是蛋白质中间体(N-外显肽-ItnN和IntC-C-外显肽)的积累，以及切下的断裂内含肽。为了解决这个问题，在本申请中，我们已经表明某些内含肽可以与某些降解信号结合，从而显著减少此类蛋白质中间体的积累。本发明集中于内含肽和降解信号的这种组合。我们还确定了有助于增加体内剪接产量的某些内含肽-降解决定子组合。

发明内容

在优选的首要方面，本发明涉及一种组合物，该组合物包括：

a.第一多核苷酸，所述第一多核苷酸编码包含断裂内含肽N片段的多肽，其中所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接的SEQ ID NO 27的CfaN、SEQ ID NO 30的CatN和SEQ ID NO 38的Gp41N、或其任何功能等效变体如SEQ ID NO 39的ConN；和

b.第二多核苷酸，所述第二多核苷酸编码包含断裂内含肽C片段的多肽，其中所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与所述要重构的蛋白质的C端片段直接连接的SEQ ID NO 28的CfaC、SEQ ID NO 31的CatC和SEQ ID NO 104的Gp41C、或其任何功能等效变体如CfaCmut(SEQ ID NO 29)或ConC(SEQ ID NO 105)；

其中所述组合物的两种多核苷酸可以一起包装在单一制剂中或分别包装在不同制剂中；

其中所述第一多核苷酸和所述第二多核苷酸分别编码所述要重构的蛋白质的所述N端片段和所述C端片段，使得当两个片段组合时，所述蛋白质的所述N端片段与所述蛋白质的所述C端片段连接，从而产生整个蛋白质；

其中所述要重构的蛋白质大于25KDa；以及

其中所述组合物进一步的特征在于：

-所述断裂内含肽N片段进一步通过肽键与降解决定子直接连接，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；和/或

-所述断裂内含肽C片段进一步通过肽键与降解决定子直接连接，其中所述降解决定子在所述内含肽C片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽C片段的所述C端通过肽键与所述要重构的蛋白质的所述C端片段直接连接。

在首选方面的优选实施方式中，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由以下组成的清单：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7。优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由以下组成的清单：DD1、DD3、PEST、SopE、L2、L9、M4或V12。更优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由以下组成的清单：SopE、L2、L9、M4或V12。

在首选方面的另一个优选实施方式中，断裂内含肽N片段是通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接的SEQ ID NO 27的CfaN，断裂内含肽C片段是通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接的SEQ ID NO 28的CfaC；其中，优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7组成的清单；更优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单；再更优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由SopE、L2、L9、M4或V12组成的清单。

在首选方面的另一个优选实施方式中，断裂内含肽N片段是通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接的SEQ ID NO 27的CfaN，断裂内含肽C片段是通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接的SEQ ID NO 29的CfaCmut；其中，优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7组成的清单；更优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单；再更优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由SopE、L2、L9、M4或V12组成的清单。

在首选方面的另一个优选实施方式中，断裂内含肽N片段是通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接的SEQ ID NO 32的NpuN，断裂内含肽C片段是通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接的SEQ ID NO 29的CfaCmut或SEQ ID NO 36的NpuCmut；其中，优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7组成的清单；更优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单；再更优选地，与断裂内含肽N片段和/或断裂内含肽C片段直接连接的降解决定子选自由SopE、L2、L9、M4或V12组成的清单。

在首选方面的另一个优选实施方式中，所述组合物的特征在于包含两个降解决定子，具体地，所述组合物的特征在于：

о断裂内含肽N片段进一步通过肽键与降解决定子直接连接，其中降解决定子在内含肽N片段和降解决定子之间有或没有接头的情况下通过内含肽的C端与内含肽N片段连接，并且其中断裂内含肽N片段的N端通过肽键与要重构的蛋白质的N端片段直接连接；以及

о断裂内含肽C片段进一步通过肽键与降解决定子直接连接，其中降解决定子在内含肽C片段和降解决定子之间有或没有接头的情况下通过内含肽的N端与内含肽C片段连接，并且其中断裂内含肽C片段的C端通过肽键与要重构的蛋白质的C端片段直接连接。

在首选方面的优选实施方式或其任何优选实施方式中，所述组合物的特征在于其包括：

a.第一多核苷酸，所述第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并可选地进一步通过肽键与选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7组成的清单中的降解决定子直接连接的SEQ ID NO 27的CfaN、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸，所述第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并可选地进一步通过肽键与选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7组成的清单中的降解决定子直接连接的SEQ ID NO 28的CfaC、或SEQ ID NO 29的CfaCmut、或SEQ ID NO 36的NpuCmut、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接。

在首选方面的优选实施方式或其任何优选实施方式中，所述组合物包括：

a.第一多核苷酸，所述第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并可选地进一步通过肽键与选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 27的CfaN、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸，所述第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并可选地进一步通过肽键与选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 28的CfaC、或SEQ ID NO 29的CfaCmut、或SEQ ID NO 36的NpuCmut、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接。

在首选方面的另一个优选实施方式或其任何优选实施方式中，所述组合物包括：

a.第一多核苷酸，所述第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并可选地进一步通过肽键与选自由SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 27的CfaN、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸，所述第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并可选地进一步通过肽键与选自由SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 28的CfaC、或SEQ ID NO 29的CfaCmut、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接。

在首选方面的另一个优选实施方式或其任何优选实施方式中，所述组合物包括：

a.第一多核苷酸，所述第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并可选地进一步通过肽键与选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7组成的清单中的降解决定子直接连接的SEQ ID NO 38的Gp41N、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸，所述第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并可选地进一步通过肽键与选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7组成的清单中的降解决定子直接连接的SEQ ID NO 104的Gp41C、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接。

在首选方面的另一个优选实施方式或其任何优选实施方式中，所述第一多核苷酸和所述第二多核苷酸分别编码ABCA4蛋白的N端片段和C端片段，使得当这两种多核苷酸被翻译成它们各自的蛋白复合物并根据本发明的方法进行组合时，所述ABCA4蛋白的所述N端片段与所述ABCA4蛋白的所述C端片段连接，从而产生完整的ABCA4蛋白。

关于上述第一和第二多核苷酸分别编码ABCA4蛋白的N端片段和C端片段的实施方式，本发明提供以下替代方案。

第一种替代方案：

a.第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并可选地进一步通过肽键与选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7组成的清单中的降解决定子直接连接的SEQ ID NO 27的CfaN、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并可选地进一步通过肽键与选自由CL1、Deg1、PESt、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7组成的清单的降解决定子直接连接的SEQ ID NO 29的CfaCmut、或SEQ ID NO 36的NpuCmut、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；

其中所述第一多核苷酸编码ABCA4蛋白的N端片段的第1-1149、1-1139、1-1176或1-1178位；并且其中所述第二多核苷酸编码ABCA4蛋白的C端片段的第1150-2273、1140-2273、1177-2273或1179-2273位，其中当所述第一多核苷酸编码第1-1149位时，所述第二多核苷酸编码第1150-2273位，其中当所述第一多核苷酸编码第1-1139位时，所述第二多核苷酸编码第1140-2273位，其中当所述第一多核苷酸编码第1-1176位时，所述二多核苷酸编码第1177-2273位，并且其中当所述第一多核苷酸编码第1-1178位时，所述第二多核酸编码第1179-2273位。

第二种替代方案：

a.第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并可选地进一步通过肽键与选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 27的CfaN、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并可选地进一步通过肽键与选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 29的CfaCmut、或SEQ ID NO 36的NpuCmut、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；

其中所述第一多核苷酸编码ABCA4蛋白的N端片段的第1-1149、1-1139、1-1176或1-1178位；并且其中所述第二多核苷酸编码ABCA4蛋白的C端片段的第1150-2273、1140-2273、1177-2273或1179-2273位，其中当所述第一多核苷酸编码第1-1149位时，所述第二多核苷酸编码第1150-2273位，其中当所述第一多核苷酸编码第1-1139位时，所述第二多核苷酸编码第1140-2273位，其中当所述第一多核苷酸编码第1-1176位时，所述第二多核苷酸编码第1177-2273位，并且其中当所述第一多核苷酸编码第1-1178位时，所述第二多核酸编码第1179-2273位。

第三种替代方案：

a.第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并可选地进一步通过肽键与选自由SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 27的CfaN、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并可选地进一步通过肽键与选自由SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 29的CfaCmut、或SEQ ID NO 36的NpuCmut、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；

其中所述第一多核苷酸编码ABCA4蛋白的N端片段的第1-1149、1-1139、1-1176或1-1178位；并且其中所述第二多核苷酸编码ABCA4蛋白的C端片段的第1150-2273、1140-2273、1177-2273或1179-2273位，其中当所述第一多核苷酸编码第1-1149位时，所述第二多核苷酸编码第1150-2273位，其中当所述第一多核苷酸编码第1-1139位时，所述第二多核苷酸编码第1140-2273位，其中当所述第一多核苷酸编码第1-1176位时，所述第二多核苷酸编码第1177-2273位，并且其中当所述第一多核苷酸编码第1-1178位时，所述第二多核酸编码第1179-2273位。

第四种替代方案：

a.第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并可选地进一步通过肽键与选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 27的CfaN、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并可选地进一步通过肽键与选自由DD1、DD3、PEST、SopE、L2、L9、M4或V12组成的清单中的降解决定子直接连接的SEQ ID NO 28的CfaC、SEQ ID NO 29的CfaCmut、或SEQ ID NO 36的NpuCmut、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；

其中所述第一多核苷酸编码ABCA4蛋白的N端片段的第1-1149位；并且其中所述第二多核苷酸编码ABCA4蛋白的C端片段的第1150-2273位，其中当所述第一多核苷酸编码第1-1149位时，所述第二多核苷酸编码第1150-2273位。

第五种替代方案：

a.第一多核苷酸编码包含断裂内含肽N片段的多肽，所述断裂内含肽N片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接、并进一步通过肽键与选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7组成的清单中的降解决定子直接连接的SEQ ID NO 38的Gp41N、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的所述N端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；以及

b.第二多核苷酸编码包含断裂内含肽C片段的多肽，所述断裂内含肽C片段选自由以下组成的清单：通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接、并进一步通过肽键与选自由CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7组成的清单中的降解决定子直接连接的SEQ ID NO 104的Gp41C、或其任何功能等效变体，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过所述内含肽的所述N端与所述内含肽C片段连接，并且其中所述断裂内含肽N片段的所述C端通过肽键与所述要重构的蛋白质的所述N端片段直接连接；

所述第一多核苷酸编码ABCA4蛋白的N端片段的第1-1095或1-1185位；以及其中所述第二多核苷酸编码ABCA4蛋白的C端片段的第1096-2273或1186-2273位，其中当所述第一多核苷酸编码第1-1095位时，所述第二多核苷酸编码第1096-2273位，并且其中当所述第一多核苷酸编码第1-1185位时，所述第二多核苷酸编码第1186-2273位。

在首选方面的进一步优选实施方式或其任何优选实施方式或替代方案中，两种多核苷酸都包含在使得多核苷酸在合适的宿主细胞中增殖或插入的载体中。优选地，所述载体是腺相关病毒(AAV)，更优选地，载体是血清型1、2、3、4、5、6、7、8或9的AAV。

在首选方面的进一步优选实施方式或其任何优选实施方式或替代方案中，本文所述的组合物用于治疗，特别是用于表1中确定的任何疾病。

本发明的另一个方面涉及用于在细胞中表达目标基因的体外或体内方法，该方法包括：

(i)使细胞与如前述方面、实施方式或替代方案中任一项所定义的的第一多核苷酸和第二多核苷酸接触，其中优选两种多核苷酸都包含在腺相关病毒(AAV)内，并且其中所述多核苷酸中的至少一个编码通过肽键与降解决定子直接连接的断裂内含肽片段，

(ii)使第一多核苷酸和第二多核苷酸表达，以产生第一融合蛋白和第二融合蛋白，以及

(iii)使第一蛋白和第二蛋白接触，使得断裂内含肽N片段与断裂内含肽C片段结合以形成内含肽中间体，并且所述内含肽中间体反应以将第一目标多肽的C端与第二目标多肽的N端共价连接。

附图说明

图1.使用内含肽以及与降解决定子结合的内含肽来重组大蛋白用于基因治疗的策略的整体方案。上图：(1)目标基因在审慎选择的位置重组断裂，所得的5'端重组融合到IntN，3’端融合到IntC，使得在蛋白质表达时，蛋白质的N端片段将与与其C端融合的IntN一起表达，而蛋白质的C端片段将与与其N端融合的IntC一起表达。(2)每个构建体都封装在单独的AAV中。将这两种AAV施用于患者，以便将它们共转导至某些细胞(3)。在共转导后，通过AAV递送的DNA将被转录成RNA并翻译成蛋白质，并且在蛋白质水平上，内含肽将进行蛋白质反式剪接反应以重构所期望的剪接产物(4)。下图：内含肽与降解信号相结合，以防止起始材料积累并消除切下的内含肽。降解决定子与N-内含肽的C端和/或C-内含肽的N端融合。

图2.上图：EGFP

图3.用EGFP-内含肽多核苷酸转染的裂解的HEK293细胞的蛋白质印迹分析。左图：用EGFP-内含肽质粒转染的细胞的蛋白质印迹分析。用全长EGFP质粒转染细胞，或用EGFP

图4.通过流式细胞术分析HEK293转染的细胞。左图：用不同构建体或构建体的组合转染的细胞的流式细胞术数据。黑色曲线对应于用EGFP的N端片段或C端片段转染的对照。蓝色是用编码全长EGFP的质粒转染的细胞的4次重复，绿色曲线对应于用EGFP

图5：在Cfa和Npu之间对ABCA4位置1150处的剪接产量进行比较。细胞用ABCA4

图6：在Cfa和Npu之间对ABCA4位置1140处的剪接产量进行比较。细胞用ABCA4

图7：对ABCA4位置1140、1150、1179和1188处的剪接产量进行比较。细胞用相应的ABCA4

图8：包括降解决定子的PTS的反应方案。目标蛋白的N端片段与IntN、接头(在这种情况下为His6标签)和降解决定子融合。蛋白质的C端与IntC融合，IntC的N端与降解决定子连接。剪接反应导致形成所期望的全长蛋白质(没有任何降解决定子与全长蛋白质融合)以及切下的内含肽(切下的内含肽各自带有降解信号)。

图9：将内含肽和降解决定子组合以重构报告蛋白EGFP。HEK293细胞用指定的构建体转染，裂解并使用抗His6标签mAb通过蛋白质印迹进行分析。结果表明，使用将CfaN或C内含肽与降解信号结合的构建体导致由起始材料(EGFP-CfaN-6H)以及内含肽(CfaN-6H)产生的任何信号消失。

图10：将内含肽和降解决定子组合以重构大蛋白ABCA4。在有和没有降解决定子的情况下，HEK293细胞用ABCA4

图11：内含肽-降解决定子组合的效果由蛋白酶体介导。在有和没有降解决定子的情况下，细胞用ABCA4

图12：使用不同内含肽或内含肽与降解决定子组合的重构产量的比较。左图：使用Cfa、Cfa-SopE或Npu对在位置1150处断裂的ABCA4通过PTS进行重构的蛋白质印迹。用ABCA4

图13：使用CfaCmut在有和没有降解信号的情况下在位置1140重构ABCA4。左图：用ABCA4

图14：内含肽和降解决定子组合以重构报告蛋白EGFP，其中使用DD1降解决定子。HEK293细胞用指定的构建体转染，裂解并使用抗His6标签mAb通过蛋白质印迹进行分析。Cfa内含肽与DD1降解决定子的组合导致消除不期望的起始材料和切下的内含肽。

图15：质谱分析。用ABCA4

图16：使用正交共有内含肽(orthogonal consensus inteins)的三段式连接。

图17：使用正交共有内含肽的三段式连接。图16所示的N、M和C构建体单独转染、成对共转染或三个片段一起共转染到HEK293中。裂解细胞并，使用针对存在于N、M和C端片段的C端的3FT标签的抗体(左图)或针对POIN片段的抗体(右图)通过蛋白质印迹分析细胞。

图18：A)分析降解决定子对本发明的断裂内含肽N片段的表达水平的影响。将具有和不具有降解决定子的构建体转染到HEK293细胞中，并通过蛋白质印迹分析表达水平。向N内含肽片段添加降解决定子导致检测到的表达蛋白质水平降低。B)分析降解决定子对本发明的断裂内含肽C片段的表达水平的影响。将具有和不具有降解决定子的构建体转染到HEK293细胞中，并通过蛋白质印迹分析表达水平。向C内含肽片段添加降解决定子导致检测到的表达蛋白质水平降低。C)示出了用N端或C端片段转染或用两种片段共转染的细胞的蛋白质印迹分析。示出了不具有降解决定子(无降解决定子)或具有代表性降解决定子(SopE、L2和L9)的构建体的结果。右图提供了与不具有降解决定子获得的产物相比，使用各指定的降解决定子获得的PTS产物量的量化。可以看出，与不具有降解决定子的构建体相比，一些降解决定子提供更高的产量。有趣的是，所有的降解决定子都有效，并且使用所有的降解决定子都可以重构在不具有降解决定子的情况下获得的产物的至少25％。一些降解决定子允许重构在不具有降解决定子的情况下获得的水平的超过50％，最后，有一些降解决定子产生100％(或更高)的重构。

图19.使用不同类型的能够检测内含肽的凝胶通过WB分析用不同降解决定子进行的PTS反应。可以看出，与其他降解决定子(例如SpoE和PEST)相比，几种降解决定子减少了切下的内含肽的量，而其他降解决定子(如L2、L9、M2、M4或DD3)则完全消除了内含肽条带。

图20.通过蛋白质印迹分析在具有不同的降解决定子组合和不具有降解决定子的情况下在位置1150的断裂处的PTS反应。在这种情况下，N片段处的降解决定子是固定的(左图L9和右图L2，标有N)，C片段处使用不同的降解决定子(L2、L9、M4、M2、V12、DD3和DD1，标有C)。

图21：在N端和C端内含肽ABCA4片段中具有或不具有降解决定子L9的情况下位点1140和11 1179处的PTS反应的WB和量化。

图22：使用gp41内含肽通过蛋白质印迹分析PTS反应。左图：使用Cfa内含肽对在位置1150处断裂的ABCA4通过PTS进行重构的蛋白质印迹和使用gp41内含肽对在位置1185处断裂的ABCA4通过PTS进行重构的蛋白质印迹。48小时后收集细胞，裂解并通过WB分析。右图：用各内含肽以及全长蛋白质获得的产物的光密度定量。

图23：体内视网膜EGFP：(A)注射了编码全长EGFP的AAV8或编码本发明的N端片段(N，EGFPN-CfaN)和C端片段(C，CfaC-EGFPC)的两种AAV的小鼠的眼底自发荧光(FAF)。(B)盐水或处理过的视网膜的免疫组织化学分析(IHC)。结果示出了光感受器中的天然EGFP荧光，指示了蛋白质剪接介导的EGFP重构。所有构建体都在GRK1启动子的控制下，并且注射了两种不同剂量的N+C构建体。BSS(平衡无菌盐水)。

具体实施方式

本发明涉及使用工程改造的或天然存在的断裂内含肽以及其与降解信号(去稳定结构域，降解决定子)的组合来重构用于基因治疗的大蛋白的方法。

本发明中描述的方法允许在体外、离体和体内重构大蛋白。当在体内使用时，该方法将允许在任何期望的组织中重构所需的大目标蛋白，所述期望的组织包括但不限于：中枢神经系统(CNS)、周围神经系统(PNS)、肌肉、肝脏、眼睛、胰腺、视网膜、肾脏、内耳、心脏、肺、血液、脾脏、皮肤。

如本文所用，术语“内含肽”是指天然存在的或人工构建的多肽序列，该多肽序列能够催化从前体蛋白切下内含肽序列并通过肽键加入侧翼序列(N-外显肽和C-外显肽)的蛋白剪接反应。内含肽的大小通常为150-550个氨基酸，也可包含归巢核酸内切酶(homingendonuclease)结构域。已知的内含肽的列表发布在http://www.inteins.com和https://inteins.biocenter.helsinki.fi/index.php上。

如本文所用的术语“断裂内含肽”是指任何这样的内含肽，其中的N端和C端氨基酸序列不通过肽键直接连接，使得N端和C端序列变成单独的片段，所述单独的片段可以非共价重新缔合或重构为对反式剪接反应(trans-splicing reaction)起作用的内含肽。

术语“肽键”是指当一个分子的羧基部分(称为羧基组分)与另一个分子的氨基部分(称为氨基组分)反应时，两个分子之间形成的共价化学键-CO-NH-，从而导致分子的释放。例如，蛋白原L-氨基酸在结合时可以形成肽键，同时释放水分子。因此，蛋白质和肽可以看作是由肽键连接在一起的氨基酸残基链。肽键是“酰胺键”或“酰胺连接”。

术语“多肽”、“肽”或“蛋白质”在本文中可互换使用以指代氨基酸的聚合物。

术语“氨基酸”是指天然存在的氨基酸和合成氨基酸，以及以类似于天然存在的氨基酸的方式起作用的氨基酸类似物和氨基酸模拟物。此外，术语“氨基酸”包括D-氨基酸和L-氨基酸(立体异构体)。

术语“天然氨基酸”或“天然存在的氨基酸”包括20种天然存在的氨基酸；这些氨基酸经常在体内进行翻译后修饰，这些氨基酸包括例如羟脯氨酸、磷酸丝氨酸和磷酸苏氨酸；和其他不常见的氨基酸，包括但不限于2-氨基己二酸、羟赖氨酸、异锁链素、正缬氨酸、正亮氨酸和鸟氨酸。

如本文所用，术语“非天然氨基酸”或“合成氨基酸”是指在“α”位处被胺基取代并且在结构上与天然氨基酸相关的羧酸或其衍生物。修饰的或不常见的氨基酸的说明性非限制性实例包括：2-氨基己二酸、3-氨基己二酸、β-丙氨酸、2-氨基丁酸、4-氨基丁酸、6-氨基己酸、2-氨基庚酸、2-氨基异丁酸、3-氨基异丁酸、2-氨基庚二酸、2,4-二氨基丁酸、锁链素、2,2'-二氨基庚二酸、2,3-二氨基丙酸、N-乙基甘氨酸、N-乙基天冬酰胺、羟基赖氨酸、别羟基赖氨酸(alio hydroxy lysine)、3-羟脯氨酸、4-羟脯氨酸、异锁链素、别异亮氨酸、N-甲基甘氨酸、N-甲基异亮氨酸、6-N-甲基-赖氨酸、N-甲基缬氨酸、正缬氨酸、正亮氨酸、鸟氨酸、对乙酰苯丙氨酸、对卤代苯丙氨酸、对丙炔氧基苯丙氨酸(p-proparglyoxyphenylalanina)、对叠氮基苯丙氨酸、对苯甲酰基苯丙氨酸等。该组还包括“天然氨基酸”的D-异构体。

如本文所用，术语“断裂内含肽N片段”或“N端断裂内含肽”或“N端内含肽片段”或“N端内含肽序列”(缩写为“IntN”)是指任何这样的内含肽，其包含对反式剪接反应起作用、即能够与功能性断裂内含肽C片段缔合以形成完整的内含肽的N端氨基酸序列，该完整的内含肽序列能够将自身从宿主蛋白中切下，催化外显肽或侧翼序列通过肽键连接，或与断裂内含肽C片段缔合时催化“N端切割”，即对外显肽和断裂内含肽N片段的N端之间的肽键的亲核攻击，从而导致所述肽键断裂。因此，lntN还包括在发生反式剪接时被剪掉的序列。lntN可以包含作为天然存在的内含肽序列的N端部分的修饰物的序列。例如，它可以包含额外的氨基酸残基和/或突变的残基，只要包含这些额外的和/或突变的残基不会使lntN在反式剪接中不起作用。优选地，包含额外的和/或突变的残基提高或增强了lntN的反式剪接活性。

如本文所用，术语“降解决定子”是指当与另一多肽重组融合时通过蛋白酶体降解途径或任何其他细胞降解机制加速其蛋白质降解的天然存在的或人工构建的多肽序列。

如本文可互换使用的，术语“断裂内含肽C片段”或“C端断裂内含肽”或“C端内含肽片段”或“C端内含肽序列”(缩写为“lntC”)是指任何这样的内含肽，其包含对反式剪接反应起作用、即在缔合时能够与功能性断裂内含肽N片段缔合以形成完整的内含肽的C端氨基酸序列，该完整的内含肽序列能够将自身从宿主蛋白中切下，催化外显肽或侧翼序列通过肽键连接，或与分裂的N内含肽缔合时催化“C端切割”，即对外显肽和断裂内含肽C片段的C端之间的肽键的亲核攻击，从而导致所述肽键断裂。因此，lntC还包括在发生反式剪接时被剪掉的序列。lntC可以包含作为天然存在的内含肽序列的C端部分的修饰物的序列。例如，它可以包含额外的氨基酸残基和/或突变的残基，只要包含这些额外的和/或突变的残基不会使lntC在反式剪接中不起作用。优选地，包含额外的和/或突变的残基提高或增强了lntC的反式剪接活性。

在此需要说明的是，在本发明的上下文中，“要重构的蛋白质的N端片段”是指蛋白质的N端片段，更具体地，是指超过25KDa、超过50KDa或超过100KDa的蛋白质片段。因此，如本文所用，术语“蛋白质的N端片段”是指包括蛋白质(其成熟或未成熟形式)的N端的长度可变的片段。在一个具体实施方式中，N端片段是包含整个蛋白质长度的小于100％、小于90％、小于80％、小于70％、小于60％、小于50％、小于40％、小于30％、小于20％、小于10％、小于5％的片段。

在此需要说明的是，在本发明的上下文中，“要重构的蛋白质的C端片段”是指蛋白质的C端片段，更具体地，是指超过25KDa、超过50KDa或超过100KDa的蛋白质片段。因此，如本文所用，术语“蛋白质的C端片段”是指包括蛋白质的C端的长度可变的片段。在一个具体实施方式中，C端片段是包含整个蛋白质长度的小于100％、小于90％、小于80％、小于70％、小于60％、小于50％、小于40％、小于30％、小于20％、小于10％、小于5％的片段。

如本文所用，术语“多核苷酸”是指由通过磷酸二酯键(或其相关结构变体或合成类似物)连接的多个核苷酸单元(脱氧核糖核苷酸或核糖核苷酸，或其相关结构变体或合成类似物)组成的聚合物。术语多核苷酸包括双链或单链基因组和cDNA、RNA、任何合成的和基因操纵的多核苷酸，以及正义多核苷酸和反义多核苷酸(尽管本发明仅公开了正义链)。这包括单链分子和双链分子，即DNA-DNA、DNA-RNA和RNA-RNA杂合体。

本发明涉及通过肽键与要重构的蛋白质的N端片段直接连接的断裂内含肽N片段。该构建体从N端到C端将具有以下一般架构：

(要重构的蛋白质的N端片段)–内含肽N。

要重构的蛋白质可以是任何其重构可以产生积极的治疗效果的大基因。可以用本发明重构的蛋白质的非限制性例子是ABCA4蛋白质，以及由下表1中列出的基因编码的蛋白质。重构这些蛋白质的目的是通过提供这些基因的正确版本来治疗与其编码基因中的突变相关的疾病。此外，要重构的蛋白质可以是治疗疾病的蛋白质或酶，但是该蛋白质或酶不一定替代突变的蛋白质或酶。例如，要重构的蛋白质可以是用于基因、碱基或先导编辑(primeediting)的CRISPR/Cas9系统。

LCA＝莱伯氏先天性黑朦症

RP＝色素性视网膜炎

ad＝常染色体显性遗传

ar＝常染色体隐性遗传

PR＝感光器

RPE＝视网膜色素上皮

ECM＝细胞外基质

因此，本发明的第一方面涉及通过肽键与要重构的蛋白质的N端片段直接连接的断裂内含肽N片段，其中，如上所反映的，IntN与N端片段的C端直接连接或通过使用接头连接(下文称为“本发明的断裂内含肽N片段”)。

本发明的第一方面的优选实施方式是指通过肽键与降解决定子直接连接的断裂内含肽N片段(下文称为“本发明的断裂内含肽N片段降解决定子”)，其中降解决定子在内含肽N片段和降解决定子之间有或没有接头的情况下通过内含肽的C端与内含肽N片段连接，并且其中断裂内含肽N片段的N端通过肽键与要重构的蛋白质的N端片段直接连接。

因此，本发明的断裂内含肽N片段降解决定子的构建体的架构从N端到C端将是：

(要重构的蛋白质的N端部分)-(内含肽-N片段)-(降解决定子)。

如果在IntN和降解决定子之间引入接头，则本发明的断裂内含肽N片段降解决定子的构建体的架构从N端到C端将是：

(要重构的蛋白质的N端部分)-(内含肽-N片段)-(接头)-(降解决定子)。

优选地，任何的本发明的断裂内含肽N片段或本发明的断裂内含肽N片段降解决定子的内含肽N可以选自下表2中列出的以下中的任意者：

其中优选地，内含肽N是SEQ ID NO 27的CfaN内含肽或其任何变体，例如SEQ IDNO 39的ConN；或SEQ ID NO 30的CatN内含肽或其任何变体；或SEQ ID NO 32的NpuN内含肽或其任何变体；或SEQ ID NO 38的Gp41N内含肽或其任何变体。

如本文所用，术语“变体”是指与特定多肽序列基本相似的多肽分子。在本发明中，我们将Cfa、Npu、Cat或Gp41内含肽的N内含肽或C内含肽的变体称为与任何其多肽序列基本相似的多肽，例如Con内含肽在本文中被理解为Cfa的变体。因此，变体可在结构和生物活性上与衍生出该变体的多肽相似。因此，变体可以指多肽序列的突变体。术语“突变体”是指这样的多肽分子，该多肽分子的序列与衍生出该多肽分子的多肽分子相比具有添加、删除、取代或以其他方式化学修饰的一个或多个氨基酸。突变体可以保留与衍生出该突变体的多肽分子基本相同的特性，或者缺乏所声称的序列的生物学活性。在特定的实施方式中，内含肽变体包括非催化性Cys残基(即N内含肽的第一个残基)已突变为丝氨酸或丙氨酸的突变体。

优选地，本发明中通常理解的术语“变体”是指混杂性(promiscuity)增加的变体，其中混杂性理解为内含肽不取决于紧邻断裂位点的残基的身份而进行蛋白质反式剪接(PTS)反应的能力。更具体地说，内含肽混杂性是指它们不取决于紧邻断裂位点(即内含肽插入目标蛋白的位点)的氨基酸的身份进行PTS反应的能力。下面示出了断裂位点的示意图：

-3 -2 -1 +1+2+3

X–X–X–内含肽N 内含肽C–X–X–X

通常，内含肽对在这些位置上的某些氨基酸具有强烈的偏好。例如，Cat内含肽(SEQ ID NO 30)偏好+1位上的Cys和-1位上的Glu(Stevens,A.J.,Sekar,G.,Gramespacher,J.A.,Cowburn,D.,&Muir,T.W.(2018)An Atypical Mechanism of SplitIntein Molecular Recognition and Folding.Journal of the American ChemicalSociety,140(37),11791–11799.http://doi.org/10.1021/jacs.8b07334)。更具混杂性的变体将是那些即使在不存在此类优选残基的情况下也能够以良好的产率进行蛋白质反式剪接反应的变体。

在具体的实施方式中，内含肽变体包括保留了内含肽的催化残基和第二壳促进剂残基(second shell accelerator residues)的突变体，并且仅非催化残基或第二壳外的残基是突变的。第二壳促进剂残基是与内含肽活性位点相邻的那些残基，它们在调节内含肽的剪接活性中起关键作用。例如，Cfa内含肽和DnaE家族的内含肽的催化残基和促进剂残基是众所周知的，并且已在本领域中描述过(Stevens,A.J.,Brown,Z.Z.,Shah,N.H.,Sekar,G.,Cowburn,D.,&Muir,T.W.(2016).Journal of the American ChemicalSociety.http://doi.org/10.1021/jacs.5b13528)。同样，已经研究和表征了多个其他内含肽家族的催化残基，包括DnaE、GyrA、GyrB、DnaB、TerL、gp41、IMPDH(Shah,N.H.,&Muir,T.W.(2014).Inteins:Nature's Gift to Protein Chemists.Chemical Science(RoyalSociety of Chemistry:2010),5(1),446–461.http://doi.org/10.1039/C3SC52951G)。还描述了识别第二壳促进剂残基的方法(Stevens et al.2016Journal of the AmericanChemical Society.http://doi.org/10.1021/jacs.5b13528)，并且所述方法可用于生成具有用于本发明的合适性质的内含肽变体(N内含肽和C内含肽)。

在具体的实施方式中，内含肽变体包括保留了内含肽的催化残基的突变体，并且该变体保留了CfaN、NpuN、CatN或gp41N内含肽的关键功能特征，包括在离液剂或高温存在下具有快速剪接速率(半衰期低于5分钟)和高活性。在更具体的实施方式中，CfaN、CatN、NpuN或gp41N内含肽的变体是任何这些序列的功能等效变体。

如本文所用，术语“功能等效变体”应理解为指所有通过修饰、插入和/或删除或一个或多个氨基酸而从序列衍生出的那些蛋白质，只要功能基本上保持或改善，特别是在断裂内含肽N片段的功能等效变体的情况下，指的是维持其活性。如本文所用，术语“活性”是指断裂内含肽N片段在与断裂内含肽C片段结合后进行蛋白质反式剪接反应的能力。

CfaN、NpuN、CatN或gp41N内含肽的功能等效变体的例子如下所示：

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CfaN序列(SEQ ID NO 27)，其中Cys28和/或Cys59突变为Ser、Thr或Ala。

CfaN序列(SEQ ID NO 27)，其中在Met1的N端包含额外的氨基酸。例如，接头、降解决定子或用于检测的标签。

CfaN序列(SEQ ID NO 27)，其中其任何残基突变为具有相似物理化学性质的残基，但必须保留的以下残基除外：Cys1、Lys70、His72、Met75、Met81。

CfaN序列(SEQ ID NO 27)，其中其任何残基突变为具有相似物理化学特性的残基，但必须保留的以下残基除外：Cys1、Asp5、Phe15、Glu24、Thr32、Lys35、Phe38、Val39、Ile44、Asn49、Ile65、Thr69、Lys70、His72、Met75、Thr77、Met81、Gly91、Lys95、Gln96、Gly99。

CfaN序列(SEQ ID NO 27)，其中保留了残基Cys1、Lys70、His72、Met75、Met81以及引入了任何以下突变或其任何组合：Asp5Glu、Phe15Leu、Glu24Lys、Thr32Ser、Lys35Asn、Phe38Asn、Val39Ile、Ile44Val、Asn49Asp、Ile65Leu、Thr77Val、Gly91Glu、Lys95Met、Gln96Arg、Gly99Asn。

CfaN序列(SEQ ID NO 27)，其中保留了残基Cys1、Lys70、His72、Met75、Met81以及引入了以下突变中的1至6个(其中1和6包含在范围内)的：Asp5Glu、Phe15Leu、Glu24Lys、Thr32Ser、Lys35Asn、Phe38Asn、Val39Ile、Ile44Val、Asn49Asp、Ile65Leu、Thr77Val、Gly91Glu、Lys95Met、Gln96Arg、Gly99Asn。

CfaN序列(SEQ ID NO 27)，其中保留残基Cys1、Asp5、Glu24、Phe38、Val39、Ile44、Lys70、His72、Met75、Met81、Gly91、Lys95、Gln96、Gly99并引入了任何以下突变或突变组合：Phe15Ala、Thr32Ser、Lys35Glu、Asn49Asp、Ile65Thr、Thr77Glu。

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-

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在具体实施方式中，内含肽N包含以下或由以下组成：分别与SEQ ID NO:27、SEQID NO 30、SEQ ID NO 32或SEQ ID NO 38在整个序列上具有至少90％序列同一性的SEQ IDNO:27、SEQ ID NO 30、SEQ ID NO 32或SEQ ID NO 38的氨基酸序列的变体。在具体实施方式中，SEQ ID NO:27、SEQ ID NO 30、SEQ ID NO 32或SEQ ID NO 38的内含肽N的变体分别与SEQ ID NO:27、SEQ ID NO 30、SEQ ID NO 32或SEQ ID NO 38在整个序列上具有至少91％、至少92％、至少93％、在至少94％、至少95％、至少96％、至少97％、至少98％或至少99％的序列同一性。

在两个或更多个氨基酸或核苷酸序列的上下文中，术语“同一性”、“相同”、“百分比同一性”或“序列同一性”是指在不考虑任何保守氨基酸取代作为序列同一性的一部分的情况下，当比较和对齐(如果需要，引入空位)以获得最大对应时，两个或更多个序列或子序列相同或具有特定百分比的相同氨基酸残基。百分比同一性可以使用序列比较软件或算法或通过目测来测量。本领域已知可用于获得氨基酸序列比对的各种算法和软件。序列比对算法的一个这样的非限制性例子是在Karlin et al.,1990,Proc.Natl.Acad.Sci.,87:2264-8中描述、如在Karlin et al.,1993,Proc.Natl.Acad.Sci.,90:5873-7进行修改以及并入N BLAST和XBLAST程序(Altschul et al.,1991,Nucleic Acids Res.,25:3389-402)的算法。在某些实施方式中，可以使用Gapped BLAST，如Altschul et al.,1997,NucleicAcids Res.25:3389-402中所描述。BLAST-2、WU-BLAST-2(Altschul et al.,1996,Methodsin Enzymology,266:460-80)、ALIGN、ALIGN-2(Genentech，加利福尼亚南旧金山)或Megalign(DNASTAR)是其他公开的可用于比对序列的可用软件程序。在某些替代实施方式中，GCG软件包中的GAP程序(其中并入了Needleman和Wunsch的算法(J.Mol.Biol.48:444-53(1970)))可用于确定两个氨基酸序列之间的百分比同一性(例如，使用Blossum 62矩阵或PAM250矩阵，空位权重为16、14、12、10、8、6或4，长度权重为1、2、3、4、5)。可选择地，在某些实施方式中，氨基酸序列之间的同一性百分比是使用Myers和Miller的算法(CABIOS,4:11 -7(1989))确定的。例如，百分比同一性可以使用ALIGN程序(2.0版本)和使用带有残基表、空位长度罚分为12、空位罚分为4的PAM120来确定。本领域技术人员可以通过特定比对软件确定最大比对的适当参数。在某些实施方式中，使用比对软件的默认参数。在某些实施方式中，将第一氨基酸序列与第二氨基酸序列的同一性百分比“X”计算为100×(Y/Z)，其中Y是在第一序列和第二序列的比对(如通过目视检查或特定的序列比对程序进行比对的)中被评分为相同匹配的氨基酸残基的数目，Z为第二序列中的残基总数。如果第二序列比第一序列长，则使用考虑了两个序列整体的全局比对，因此每个序列中的所有字母和空值都必须进行比对。在这种情况下，可以使用与上述相同的公式，但使用第一和第二序列重叠的区域的长度作为Z值，所述区域的长度与第一序列的长度基本相同。

作为非限制性实例，在一些实施方式中，可以使用Bestfit程序(Wisconsin序列分析包，Version 8for Unix，Genetics Computer Group，University Research Park，575Science Drive，Madison，Wl 5371 1)来确定任何特定多肽是否与参考序列具有一定百分比的序列同一性(例如，至少80％同一性、至少85％同一性、至少90％同一性，并且，至少95％、96％、97％、98％或99％同一性)。Bestfit使用Smith和Waterman的局部同源算法(Advances in Applied Mathematics 2:482-9(1981))来寻找两个序列之间的最佳同源片段。当使用Bestfit或任何其他序列比对程序来确定特定序列是否与根据本发明的参考序列具有例如95％的同一性时，设置参数使得在参考氨基酸序列的全长上计算同一性百分比并且允许同源性中的空位最多达参考序列中核苷酸总数的5％。

断裂内含肽N片段和降解决定子之间的术语“接头”是指连接内含肽N片段的C端和降解决定子序列的N端的氨基酸序列。接头将优选地是1至100个氨基酸、或1至5、或1至10、或1至50、或1至25个氨基酸的多肽。接头可以是富含Gly的肽或富含Gly-Ser的肽。富含Gly-Ser的肽的例子是序列GGS，或具有通式(GGS)n的该接头的聚合物，其中n可以为1到10。接头的通式为((G)nS)y，其中n为1到5，y为1到10。此外，表位标签可以用作接头，例如六组氨酸(His6，序列：GHHHHHHG(SEQ ID NO 84)标签，或三重flag标签(3FT，序列：DYKDHDGDYKDHDIDYKDDDDK(SEQ ID NO 103))。

在另一个具体实施方式中，通过肽键与断裂内含肽N片段直接连接的降解决定子选自下表3中列出的那些中的任意者：

降解决定子的其他非限制性实例包括与DHFR(脱氢叶酸还原酶)、FKBP(FK506结合蛋白)、FRB(FKBP-雷帕霉素结合蛋白)和PDE5(5型磷酸二酯酶)相关的多肽。以及内质网相关降解(ERAD)途径的降解信号。

在又一个优选的实施方式中，降解决定子是少于75个氨基酸的多肽，一旦该多肽与本发明的内含肽N片段融合，它就会诱导该内含肽N片段降解。

在又一个优选的实施方式中，降解决定子是这样的多肽，当其与本发明的断裂内含肽N端片段或C端片段融合时，则导致所述片段的表达水平降低超过10％，或超过20％，或超过30％，或超过50％，或超过75％，或超过90％。重要的是，在本发明的片段中掺入降解决定子不会显著影响由本发明的断裂内含肽N端片段降解决定子和C端片段降解决定子之间的蛋白质反式剪接产生的重构蛋白质的产量。为了该优选实施方式的目的，表达水平如图18中所述进行测量，即在类似于图18中描述的条件下，将具有降解决定子的片段的表达水平与不具有降解决定子的片段的表达水平进行比较，前者需要比后者低至少10％，或至少20％，或至少30％，或至少50％，或至少75％，或至少90％。

在又一个优选实施方式中，本发明的断裂内含肽N片段降解决定子的N内含肽和降解决定子的优选组合选自由以下组成的清单：

-CfaN或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7。优选地，CfaN或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4；或者-Gp41N或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7。优选地，Gp41N或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4。

在图18A中示出了一些代表性的降解决定子在与分裂的N内含肽片段融合、特别是与CfaN内含肽融合时具有的效果。添加降解决定子(SopE、L2、L9、M4、V12)导致与降解决定子融合的含有N内含肽的融合蛋白的可检测表达水平显著降低。有趣的是，如实施例和图18、20和21中所述，包含一些优选的降解决定子(SopE、L2、L9、M4或V12)不会对剪接产物形成的产量产生负面影响。现有技术并未预料到这一结果，因为当降低蛋白质反式剪接反应的起始材料水平时(如通过添加降解决定子所实现的)，人们预计蛋白质剪接产量会降低。

在又一个优选实施方式中，断裂内含肽N片段和降解决定子的优选组合选自由以下组成的清单：CfaN内含肽与任何以下降解决定子DD1、DD2、DD3、SopE、L2、L9、M4、V12、DD4、DD5、DD6或DD7的组合。

因此，我们观察到，当CfaN内含肽与上述任何降解决定子(尤其是以下降解决定子：DD1、DD2、DD3、SopE、L2、L9、M4、V12、DD4、DD5、DD6或DD7)组合时，检测到N端内含肽片段表达水平降低(图9、18A)。重要的是，这不会影响蛋白质反式剪接反应的产量(图9、18C、20或21)。这一结果是出乎意料的，根据现有技术，通常情况下，蛋白质反式剪接反应中涉及的两个内含肽片段之一的产量、表达或稳定性的降低会导致形成的剪接产物量的总体减少。

我们已经观察到膜蛋白如ABCA4(图18、20和21)以及细胞溶质可溶性蛋白(如EGFP)的这种效果，这说明了效果的普遍性。在图9中，可以观察到向EGFPN-CfaN中添加降解决定子如何导致对应于该蛋白质的条带完全消失。然而，在添加CfaC-EGFPC片段(具有或不具有降解决定子)后，形成的全长EGFP产物的产量与不使用降解决定子的情况相同。该示例说明添加一个或甚至两个降解决定子(每个片段上一个)都不会对目标蛋白的产量产生负面影响。

在另一方面，本发明涉及任何以下复合物：本发明的断裂内含肽N片段或本发明的断裂内含肽N片段降解决定子，下文称为本发明的第一复合物，其中这些复合物中的每个复合物都包括：

(i)要重构的目标蛋白的N端片段，和

(ii)断裂内含肽N片段或如以上部分中所定义的通过肽键、可选地通过接头与降解决定子直接连接的断裂内含肽N片段(本发明的断裂内含肽N片段降解决定子)；

其中复合物可选地包含(i)和(ii)之间的接头，并且

其中

-目标蛋白的N端片段通过酰胺键与断裂内含肽N片段的N端连接，或者

-如果复合物包含接头，则目标蛋白的N端片段通过酰胺键与接头结合和/或接头通过酰胺键与断裂内含肽N片段的N端结合。

可用于任何上述复合物的目标蛋白的说明性非限制性实例是上表1中所示的那些。其他目标蛋白可以选自抗体、抗体片段，包括Fc结构域、scFv、纳米抗体、双特异性抗体、蛋白质，并且优选地，大于25KDa、50KDa、100KDa的任何蛋白质。

如已经指出的，可选地，目标蛋白和断裂内含肽N片段可以通过接头连接，因此接头位于目标蛋白和N内含肽之间。接头的性质将取决于目标蛋白的性质。在具体实施方式中，接头是肽。在具体实施方式中，接头是长度为1、2、3、4、5、10、20、50、100或更多个氨基酸残基的肽；具体可以是1-3个氨基酸残基。优选地，接头的N端与目标蛋白的C端连接并且接头的C端通过肽键与N-内含肽的N端连接。

在另一个具体实施方式中，复合物在目标蛋白的N端片段和断裂内含肽N片段之间不包含接头。在该特定的实施方式中，目标蛋白通过酰胺键与断裂内含肽N片段的N端连接。

在另一个具体实施方式中，如果复合物包含接头，则复合物是融合蛋白。术语“融合蛋白”是本领域熟知的，是指人工设计的单个多肽链，其包含来自不同来源的天然和/或人工的两个或更多个序列。根据定义，融合蛋白从未在自然界中被发现。

在另一个方面，本发明涉及通过肽键与要重构的蛋白质的C端片段直接连接的断裂内含肽C片段(下文称为“本发明的断裂内含肽C片段”)。

更优选地，本发明涉及通过肽键与降解决定子直接连接的断裂内含肽C片段(下文称为“本发明的断裂内含肽C片段降解决定子”)，其中降解决定子在内含肽C片段和降解决定子之间有或没有接头的情况下通过内含肽的N端与内含肽C片段连接，并且其中断裂内含肽C片段的C端通过肽键与要重构的蛋白质的C端片段的N端直接连接。

因此，本发明的断裂内含肽C片段降解决定子的构建体的架构从N端到C端将是：

(降解决定子)-(内含肽-C片段)-(要重构的蛋白质的C端部分)

如果在降解决定子和内含肽C之间引入接头，则本发明的断裂内含肽C片段降解决定子的构建体的架构从N端到C端将是：

(降解决定子)-(接头)-(内含肽-C片段)-(要重构的蛋白质的C端部分)

优选地，任何的本发明的断裂内含肽C片段或本发明的断裂内含肽C片段降解决定子的内含肽C选自下表4中列出的以下中的任意者：

其中优选地，内含肽C是：SEQ ID NO 28的CfaC内含肽或其任何变体，例如CfaCmut(SEQ ID NO29)或ConC(SEQ ID NO 105)；或SEQ ID NO 31的CatC内含肽或其任何变体；或NpuC内含肽(SEQ ID NO 33)或其任何变体，例如NpuCmut(SEQ ID NO 36)；或SEQ ID NO104的GP41C内含肽或其任何变体；或上述定义的混杂性增加的任何变体。

术语变体与针对断裂内含肽N片段的定义相同。内含肽C的变体包括没有N端甲硫氨酸残基的SEQ ID NO 28、29、31、33、36、104和105。

在特定的实施方式中，混杂性增加的CfaC、NpuC或ConC变体在第21、22和23位包含氨基酸Gly、Glu和Pro，如Stevens et al.Proc.Natl.Acad.Sci.2017所述。

在特定的实施方式中，Cfac、NpuC CatC或gp41c内含肽的功能等效变体的实例如下所示：

-

CfaC(SEQ ID NO 28)序列，其中去除了Met1。

CfaC(SEQ ID NO 28)序列，其中在Met1的N端包含额外的氨基酸的。例如，接头、降解决定子或用于检测的标签。

CfaC(SEQ ID NO 28)序列，其中CfaC的任何残基突变为具有相似物理化学性质的残基，但应保留的以下残基除外：Asp17、His24、Asn36。

CfaC(SEQ ID NO 28)序列，其中CfaC的任何残基突变为具有相似物理化学性质的残基，但应保留的以下残基除外：Asp17、Asp23、His24、Ser35、Asn36。

CfaC(SEQ ID NO 28)序列，其中CfaC的任何残基突变为具有相似物理化学性质的残基，但应保留的以下残基除外：Asp17、Asp23、His24、Ser35、Asn36的。

CfaC(SEQ ID NO 28)序列，其中保留残基Asp17、Asp23、His24、Ser35、Asn36，并且氨基酸Val2、Ile5、Ser6、Ser9、Lys22、Leu27、Leu32，Val33中的任何氨基酸发生突变。

CfaC(SEQ ID NO 28)序列，其中残基Glu21Gly、Lys22Glu和Asp23Pro发生突变。该变体对应于CfaCmut内含肽(SEQ ID NO 29)。

CfaC(SEQ ID NO 28)序列，其中CfaC的任何残基突变为具有相似物理化学性质的残基，但残基Asp17、Gly21、Glu22、Pro23、His24、Asn36应位于在指定位置上的。

的CfaC(SEQ ID NO 28)序列，其中残基Asp17、Gly21、Glu22、Pro23、His24、Ser35、Asn36应位于指定位置并且以下氨基酸中的任何氨基酸发生突变：Val2、Ile5、Ser9、Leu27、Leu32、Val33。

CfaC(SEQ ID NO 28)或CfaCmut(SEQ ID NO 29)序列，其中引入了以下突变中的1至5个(其中1和5在范围内)：Val2Ile、Ile5Ala、Ser6Thr、Ser9Tyr、Thr12Lys、Leu27Ala、Leu32Phe、Val33Ile。

-

NpuC(SEQ ID NO 33)序列，其中去除了Met1。

NpuC(SEQ ID NO 33)序列，其中在Met1的N端包含额外的氨基酸。例如，接头、降解决定子或用于检测的标签。

NpuC(SEQ ID NO 33)序列，其中NpuC的任何残基可突变为具有相似物理化学性质的残基，但应保留的残基Asp17、His24、Asn36除外。

NpuC(SEQ ID NO 33)序列，其中NpuC的任何残基可突变为具有相似物理化学性质的残基，但需要保留残基Asp17、His24、Ser35、Asn36除外。

NpuC(SEQ ID NO 33)序列，其中保留残基Asp17、His24、Ser35、Asn36并且氨基酸Val2、Ile5、Ser9、Leu27、Leu32，Val33中的任何氨基酸发生突变。

NpuC(SEQ ID NO 33)序列，其中保留残基Asp17、His24、Ser35、Asn36并且掺入了以下突变Ile2Val,Ala5Ile,Tyr9Ser、Ala27Leu、Phe32Leu、Ile33Val中的任何突变。

NpuC(SEQ ID NO 33)序列，其中残基Glu21Gly、Arg22Glu、Asp23Pro发生突变以增加Npu的混杂性。

-

Gp41C(SEQ ID NO 104)序列，其中去除了Met1。

Gp41C(SEQ ID NO 104)序列，其中在Met1的N端包含额外的氨基酸。例如，接头、降解决定子或用于检测的标签。

Gp41C(SEQ ID NO 104)序列，其中Gp41C的任何残基可突变为具有相似物理化学性质的残基，但需要保留的残基His26、His36、Asn37除外。

在具体实施方式中，内含肽C包含以下或由以下组成：分别与SEQ ID NO：28、SEQID NO：31、SEQ ID NO：33或SEQ ID NO：104在整个序列上具有至少90％序列同一性的SEQID NO：28、SEQ ID NO：31、SEQ ID NO：33或SEQ ID NO：104的氨基酸序列的变体。在具体实施方式中，SEQ ID NO：28、SEQ ID NO：31、SEQ ID NO：33或SEQ ID NO：104的内含肽C的变体分别与SEQ ID NO：28、SEQ ID NO：31、SEQ ID NO：33或SEQ ID NO：104在整个序列上具有至少91％、至少92％、至少93％、在至少94％、至少95％、至少96％、至少97％、至少98％或至少99％的序列同一性。

在另一个具体实施方式中，通过肽键与断裂内含肽C片段直接连接的降解决定子选自表3中列出的那些中的任意者：可用于该部分的目标蛋白的说明性非限制性实例是上表1中所示的那些。

在又一个优选实施方式中，本发明的断裂内含肽C片段降解决定子的C内含肽和降解决定子的优选组合选自由以下组成的清单：

-CfaC或其任何功能等效变体(如CfaCmut)与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7。优选地，CfaC或其任何功能等效变体(如CfaCmut)与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12、DD4、DD5、DD6或DD7；或者

-Gp41C或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7。优选地，Gp41C或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4。

在图18B中，可以观察到将降解决定子与C端构建体融合的效果。正如对于N端降解决定子所观察到的，降解决定子与C端构建体的融合降低了构建体的表达水平，但当使用优选的组合时，没有观察到对剪接产量的影响。以下降解决定子与CfaC配合得特别好：DD1、DD2、DD3、SopE、L2、L9、M4、V12、DD4、DD5、DD6或DD7。正如对N端片段所观察到的，将降解决定子掺入本发明的C端片段导致其表达降低(如通过WB检测的)，但同样有趣的是，对PTS产量没有负面影响，即观察到形成了所需产物。已在若干种蛋白质中观察到该结果，包括这里示出的EGFP和ABCA4的示例。

在另一方面，本发明涉及任何以下复合物：本发明的断裂内含肽C片段或本发明的断裂内含肽C片段降解决定子，下文称为本发明的第二复合物，其中这些复合物中的每个复合物都包括：

(i)目标蛋白的C端片段，和

(ii)断裂内含肽C片段或如以上部分中所定义的通过肽键、可选地通过接头与降解决定子直接连接的断裂内含肽C片段(本发明的断裂内含肽C片段降解决定子)；

其中复合物可选地包含(i)和(ii)之间的接头，并且

其中

-目标蛋白的C端片段通过酰胺键与断裂内含肽C片段的C端连接，或者

-如果复合物包含接头，则目标蛋白的C端片段通过酰胺键与接头结合和/或接头通过酰胺键与断裂内含肽C片段的C端结合。

可用于本发明的目标蛋白的说明性非限制性实例是上表1中列出的那些。其他目标蛋白的例子可以选自抗体、抗体片段，包括Fc结构域、scFv、纳米抗体、双特异性抗体、蛋白质，并且优选地，大于25KDa、50KDa、100KDa的任何蛋白质。

先前已经结合本发明的第一复合物对术语“目标蛋白”和“接头”进行了定义。本发明的第一复合物的目标蛋白和接头的所有具体实施方式完全适用于本发明的第二复合物。

在具体实施方式中，复合物在目标化合物和断裂内含肽C片段之间不包含接头。在该特定的实施方式中，目标化合物通过酰胺键与断裂内含肽C片段的C端连接。

在具体实施方式中，复合物在目标化合物和断裂内含肽C片段之间包含接头。在该特定的实施方式中，取决于目标化合物和接头的化学性质，目标化合物可以通过任何合适的方式与接头结合。在该特定的实施方式中，接头通过酰胺键与断裂内含肽C片段的C端结合。在另一个具体实施方式中，目标化合物通过酰胺键与接头结合，在这种情况下接头可以通过任何合适的方式与断裂内含肽C片段的C端结合。在另一个具体实施方式中，目标化合物通过酰胺键与接头结合并且接头通过酰胺键与断裂内含肽C片段的C端结合。

在具体实施方式中，如果复合物包含接头，则接头是肽接头。在该特定的实施方式中，复合物是融合蛋白。

在另一个方面，本发明涉及一种组合物，以下称为本发明的第一组合物，其包含本发明的第一复合物和/或第二复合物。

术语“组合物”旨在涵盖包含特定组分的产物，以及直接或间接由特定量的特定组分组合而成的任何产物。组合物的组分可以一起包装在单一制剂中或分别包装在不同制剂中；因此，在实施方式中，本发明的第一复合物与本发明的第二复合物一起包装在单一制剂中。在另一个实施方式中，本发明的第一复合物和本发明的第二复合物是分开包装的。

在特别优选的实施方式中，第一复合物和第二多复合物分别包含相同蛋白质的N端片段和C端片段，使得当根据本发明的方法组合两种复合物时，蛋白质的所述N端片段与所述蛋白质的所述C端片段连接，从而产生整个蛋白质。

在另一方面，本发明涉及编码本发明的第一复合物或第二复合物的多核苷酸。

优选地，在优选的实施方式中，本发明涉及两种多核苷酸，其中这些多核苷酸中的一种编码本发明的第一复合物(下文称为本发明的第一多核苷酸)，和另一种编码本发明的第二复合物(在下文中称为本发明的第二多核苷酸)，其中本发明的第一和第二多核苷酸分别编码同一蛋白质的N端片段和C端片段，使得当两种多核苷酸都翻译成它们各自的蛋白质复合物时和/或当根据本发明的方法组合时，蛋白质的N端片段与蛋白质的C端片段连接，从而产生要重构的完整蛋白质。值得注意的是，本发明的第一和第二多核苷酸各自优选地编码同一内含肽的断裂内含肽，也就是说，N内含肽和C内含肽是同源对(cognate pairs)(参见Shah et al.Journal of the American Chemical Society 2012

在优选的实施方式中，本发明涉及上述所指的两种多核苷酸，其中这些多核苷酸中的一种编码本发明的断裂内含肽N片段降解决定子(下文称为本发明的编码降解决定子的第一多核苷酸)，另一种编码本发明的断裂内含肽C片段降解决定子(下文称为本发明的编码降解决定子的第二多核苷酸)，其中本发明的编码降解决定子的第一多核苷酸和本发明的编码降解决定子的第二多核苷酸分别编码同一蛋白质的N端片段和C端片段，使得当两种多核苷酸都翻译成它们各自的蛋白质复合物时复合物并根据本发明的方法组合时，蛋白质的N端片段与蛋白质的C端片段连接，从而产生完整蛋白质。与前述实施方式一样，值得注意的是，本发明的编码降解决定子的第一多核苷酸和本发明的编码降解决定子的第二多核苷酸各自优选编码同一内含肽的断裂内含肽。无论如何，如上所反映的，如本文所用，各多核苷酸必须编码断裂内含肽，使得一旦翻译成蛋白质，N端序列和C端序列会变成可非共价重新缔合或重构为对反式剪接反应起作用的内含肽的单独的片段。

在又一个优选实施方式中，本发明涉及使用正交断裂内含肽对的三段式连接策略。在该方法中，存在三种多核苷酸，其中第一多核苷酸编码POI

在此要注意的是，对于那些涉及两个多核苷酸的实施方式，第一多核苷酸(或特别是本发明的编码降解决定子的第一多核苷酸)可以是任何本发明的断裂内含肽N片段或任何本发明的断裂内含肽-N降解决定子片段。特别地，本发明的第一多核苷酸(或特别是本发明的编码降解决定子的第一多核苷酸)可以选自由以下组成的清单中的任意者：SEQ ID NO1、3、5、8、10、12、14，17、19、21、23、24、25、66、68、70、72、74、76和78。更优选地，本发明的编码降解决定子的第一多核苷酸编码内含肽和降解决定子的任何以下特定组合：

-CfaN或其任何功能等效变体(如ConN)与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7。优选地，CfaN或其任何功能等效变体(如ConN)与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4；或者

-Gp41N或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7。优选地，Gp41N或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4。

在此还要注意的是，对于那些涉及两个多核苷酸的实施方式，第二多核苷酸(或特别是本发明的编码降解决定子的第二多核苷酸)可以是本发明的任何断裂内含肽C片段或本发明的任何断裂内含肽-C降解决定子片段。特别地，本发明的第二多核苷酸(或特别是本发明的编码降解决定子的第二多核苷酸)可以选自由以下组成的清单中的任意者：SEQ IDNO 2、4、6、9、11、13、15、18、20、22、26、67、69、71、73、75、77和79。更优选地，本发明的编码降解决定子的第二多核苷酸编码内含肽和降解决定子的任何以下特定组合：

-CfaC或其任何功能等效变体(如CfaCmut或ConC)与以下任何降解决定子组合：CL1、Deg1、PESTt、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7。优选地，CfaC或其任何功能等效变体(如CfaCmut)与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12、DD4、DD5、DD6或DD7；或者

-Gp41C或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7。优选地，Gp41C或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4。

在优选的实施方式中，本发明的编码降解决定子的第一多核苷酸和本发明的编码降解决定子的第二多核苷酸分别编码ABCA4蛋白的N端片段和C端片段，使得当这两种多核苷酸被翻译成它们各自的蛋白复合物并根据本发明的方法组合时，ABCA4蛋白的N端片段与ABCA4蛋白的C端片段连接，从而产生完整的ABCA4蛋白。更优选地，本发明的编码降解决定子的第一多核苷酸编码ABCA4蛋白的N端片段的第1-1149、1-1139、1-1178或1-1187位以及内含肽和降解决定子的任何以下特定组合：

-CfaN或其任何功能等效变体(如ConN)与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7。优选地，CfaN或其任何功能等效变体(如ConN)与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4；以及

本发明的编码降解决定子的第二多核苷酸编码ABCA4蛋白的C端片段的第1150-2273、1140-2273、1179-2273或1188-2273位以及内含肽和降解决定子的任何以下特定组合：

-CfaC或其任何功能等效变体(如CfaCmut或ConC)与以下任何降解决定子组合：CL1、Deg1、PESt、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6和DD7。优选地，CfaC或其任何功能等效变体(如CfaCmut)与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12、DD4、DD5、DD6或DD7。

值得注意的是，由于ABCA4蛋白的N端片段必须与ABCA4蛋白的C端片段连接，从而产生完整的ABCA4蛋白，当本发明的编码降解决定子的第一多核苷酸编码第1-1149位时，则本发明的编码降解决定子的第二多核苷酸编码ABCA4蛋白的C端片段的第1150-2273位。其余位置也一样。

使用上述优选的组合，可以观察到，可以获得目标蛋白(在这些实施例中为ABCA4)的有效重构，同时显著降低起始N端片段和C端片段的水平。在构建体之一或这两个构建体中包含至少一个降解决定子，由于降解加速，导致N端片段和/或C端片段的表达减少。根据现有技术水平，预计起始材料水平的这种降低将导致剪接产物的减少。然而，我们观察到相反的表现，全长剪接产物的产量不受影响，并且在某些情况下甚至可以提高。此外，我们观察到，通过使用这里描述的方法重构蛋白质，我们可以生成功能性蛋白质，如使用ATP酶活性测定所测定的。我们还观察到目标蛋白在小鼠视网膜体内的有效重构。

在另一个优选的实施方式中，本发明的编码降解决定子的第一多核苷酸编码ABCA4蛋白的N端片段的第1-1095或1-1185位以及内含肽和降解决定子的任何以下特定组合：

-Gp41N或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7。优选地，Gp41N或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4；以及

本发明的编码降解决定子的第二多核苷酸编码ABCA4蛋白的C端片段的第1096-2273或1186-2273位以及内含肽和降解决定子的任何以下特定组合：

-Gp41C或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、PEST、DD1、DD2、DD3、M1、M2、SopE、SopE-1-78、SopE-15-78、SopE-15-50、L2、L6、L9、L10、L11、L12、L15、L16、M3、M4、M5、V12、DD4、DD5、DD6或DD7。优选地，Gp41C或其任何功能等效变体与以下任何降解决定子组合：CL1、Deg1、DD1、DD2、DD3、SopE、L2、L9、M4、V12或DD4。

与前面的情况一样，值得注意的是，由于ABCA4蛋白的N端片段必须与ABCA4蛋白的C端片段连接，从而产生完整的ABCA4蛋白，当本发明的编码降解决定子的第一多核苷酸编码第1-1095位时，则本发明的编码降解决定子的第二多核苷酸编码ABCA4蛋白的C端片段的第1096-2273位。其余位置也一样。

编码上述序列的核苷酸序列(分别对于本发明的第一和第二多核苷酸或编码本发明的降解决定子的第一和第二多核苷酸)的其他优选组合选自以下任何对：1和2；3和4；5和6；66和67；68和69；70和71；72和73；74和75；76和77；以及78和79。

应注意，术语“本发明的第一和第二多核苷酸”尤其涵盖“编码本发明的降解决定子的第一和第二多核苷酸”，因此在下文中因此，我们只将本发明的第一和第二多核苷酸作为已经涵盖上述所有多核苷酸替代物的术语。

优选地，本发明涉及一种组合物，在下文中称为本发明的第二组合物，其包含本发明的第一和/或第二多核苷酸或编码本发明的降解决定子的第一和/或第二多核苷酸，和/或单独或联合地包含使用正交分裂内含肽对的三段连接体系的POI-内含肽片段中的每个。在此特定上下文中，术语“组合物”旨在涵盖含有特定组分的产物。组合物的组分可以一起包在单一制剂中或分别包在不同制剂中。因此，在实施方式中，本发明的第一多核苷酸与本发明的第二多核苷酸一起包装在单一制剂中。在另一个实施方式中，本发明的第一多核苷酸和本发明的第二多核苷酸是分开包装的。更优选地，本发明的第一第二多核苷酸分别编码要重构的蛋白质的N端片段和C端片段，使得当两种复合物根据本发明的方法组合时，蛋白质的N端片段与蛋白质的C端片段连接，从而产生整个要重构的蛋白质。

可以发现本发明的所有上述多核苷酸本身是分离的，或形成允许所述多核苷酸在合适的宿主细胞中递送和/或增殖的载体的一部分。因此，在另一个方面，本发明涉及一种载体，该载体包含如上所述的本发明的任何上述多核苷酸。

适合插入所述多核苷酸的载体是：衍生自原核生物中的表达载体的载体，例如pUC18、pUC19、Bluescript及其衍生物、mpl8、mpl9、pBR322、pMB9、ColEl、pCR1、RP4、噬菌体和“穿梭”载体(例如pSA3和pAT28)；酵母中的表达载体，例如2微米质粒、整合质粒、YEP载体、着丝粒质粒等类型的载体；昆虫细胞中的表达载体，例如pAC系列和pVL的载体；植物中的表达载体，如pIBI、pEarleyGate、pAVA、pCAMBIA、pGSA、pGWB、pMDC、pMY、pORE系列等；以及真核细胞中的表达载体，包括适用于使用任何市售杆状病毒系统转染昆虫细胞的杆状病毒。用于真核细胞的载体优选包括病毒载体(腺病毒、腺相关病毒(AAV)、与腺病毒相关的病毒(例如逆转录病毒，特别是慢病毒))以及非病毒载体，例如pSilencer4.1-CMV(Ambion)，pcDNA3、pcDNA3.1/hyg、pHMCV/Zeo、pCR3.1、pEFI/His、pIND/GS、pRc/HCMV2、pSV40/Zeo2、pTRACER-HCMV、pUB6/V5-His、pVAX1、pZeoSV2、pCI、pSVL和PKSV-10、pBPV-1、pML2d和pTDT1。优选地，载体是腺相关病毒(AAV)。优选地，载体是血清型1、2、3、4、5、6、7、8或9的AAV。二聚型或自身互补AAV载体(scAAV)也可用于插入所述多核苷酸。

因此，本发明优选地涉及开发AAV作为基因治疗载体。优选地，这些载体通过从载体的DNA中除去rep和cap而消除了它们的整合能力。所需基因(多核苷酸)与驱动基因转录的启动子一起插入在末端反向重复(ITR)之间，该末端反向重复(ITR)有助于在单链载体DNA被宿主细胞DNA聚合酶复合物转化为双链DNA之后，在细胞核中形成串联体。基于AAV的基因治疗载体在宿主细胞核中形成游离型串联体。在非分裂细胞中，这些串联体在宿主细胞的生命周期内保持完整。在分裂细胞中，AAV DNA通过细胞分裂而丢失，因为游离型DNA不与宿主细胞DNA一起复制。AAV DNA随机整合到宿主基因组中是可检测的，但发生的频率非常低。

所需基因(多核苷酸)可以与在要表达构建体的预期宿主细胞中起作用的调节元件组合。本领域普通技术人员可以选择调节元件用于合适的宿主细胞，例如哺乳动物或人宿主细胞。调节元件包括例如启动子、转录终止序列、翻译终止序列、增强子、信号肽和聚腺苷酸化元件。本发明的多核苷酸可以与启动子序列可操作地连接。考虑用于本发明的启动子包括但不限于：天然基因启动子、巨细胞病毒(CMV)启动子(KF853603.1，bp 149-735)、嵌合CMV/鸡β-肌动蛋白启动子(CBA)和截短形式的CBA(smCBA)启动子(US8298818和AAVGC 1处理的产后鸟苷酸环化酶-1敲除小鼠视网膜的视锥中的光驱动锥阻素易位(Light-DrivenCone Arrestin Translocation in Cones of Postnatal Guanylate Cyclase-1KnockoutMouse Retina Treated with AAVGC 1))、视紫红质启动子(NG 009115，bp 4205-5010)、感光器间类视黄醇结合蛋白肽(IRBP)启动子(NG_029718.1，bp 4777-5011)、卵黄样黄斑营养不良2(VMD2)启动子(NG 009033.1、bp 4870-5470)、PR特异性人G蛋白偶联受体激酶1(hGRK1；AY327580.1，bp l793-2087或bp 1793-1991)(Haire等人，2006；美国专利号8,298,818)、近端鼠视紫红质启动子(MOPS)。然而，可以使用本领域已知的任何合适的启动子。在具体的实施方式中，启动子是CMV或hGRK1启动子。在一个实施方式中，启动子是在一种或一组组织中显示选择性活性，但在其他组织中活性较低或无活性的组织特异性启动子。在一个实施方式中，启动子是感光器特异性启动子。在其他实施方式中，启动子是视锥细胞特异性和/或视杆细胞特异性启动子。

优选的启动子是CMV、GRK1、CBA和IRBP启动子。还优选的启动子是使来自各种启动子的调节元件组合而成的杂合启动子(例如使来自CMV启动子的增强子、CBA启动子和Sv40嵌合内含子组合而成的嵌合CBA启动子，本文称为CBA杂合启动子。

AAV还表现出非常低的免疫原性，似乎仅限于产生中和抗体，而它们不会诱导明确定义的细胞毒性反应。这一特征以及感染静止期细胞的能力表明它们在作为用于人类基因治疗的载体具有优于腺病毒的优势。

AAV基因组、转录组和蛋白质组：AAV基因组由正义或反义的单链脱氧核糖核酸(ssDNA)构成，其长度约为4.7千碱基。基因组包括位于DNA链两端的末端反向重复(ITR)和两个开放阅读框(ORF)：rep和cap。前者由编码AAV生命周期所需的Rep蛋白的四个重叠基因组成，后者包含衣壳蛋白VP1、VP2和VP3的重叠核苷酸序列，衣壳蛋白VP1、VP2和VP3相互作用形成二十面体对称的衣壳。

每个末端反向重复(ITR)序列包含145个碱基。它们之所以如此命名是因为它们的对称性，这被证明是有效扩增AAV基因组所必需的。这些序列的另一个特性是它们形成发夹结构的能力，这有助于所谓的自引发，该自引发使得第二条DNA链不依赖于引发酶而进行合成。还表明ITR是将AAV DNA整合到宿主细胞基因组(人类第19条染色体)并从中拯救、以及AAV DNA的有效衣壳化连同产生完全组装的脱氧核糖核酸酶抗性AAV颗粒所必需的。

在基因治疗方面，ITR似乎是邻接于治疗基因的以顺式方式需要的唯一序列：结构(cap)基因和包装(rep)基因可以反式递送。在这种假设下，建立了许多有效产生含有报告基因或治疗基因的重组AAV(rAAV)载体的方法。然而，也有发表称ITR并不是用于有效复制和衣壳化的以顺式方式需要的唯一元件。一些研究小组已经在rep基因的编码序列中确定了被称为顺式作用Rep依赖元件(CARE)的序列。当以顺式方式存在时，显示出CARE增加了复制和衣壳化。

截至2006年，已经描述了11种AAV血清型。所有已知的血清型都可以感染来自多种不同组织类型的细胞。组织特异性由衣壳血清型决定，对AAV载体进行假型化(pseudotyping)以改变其嗜性(tropism)范围可能对其在治疗中的使用很重要。在本发明中，AVV血清型1、2、3、4、5、6、7、8或9的ITR是优选的。

血清型2(AAV2)迄今为止受到了最广泛的检验。AAV2呈现出对骨骼肌、神经元、血管平滑肌细胞和肝细胞的天然嗜性。

载体还可以包含报告基因或标记基因，所述报告基因或标记基因使得能够识别那些在与载体接触后掺入了载体的细胞。

本发明上下文中有用的报告基因包括lacZ、萤光素酶、胸苷激酶、GFP等。本发明上下文中有用的标记基因包括：例如，新霉素抗性基因，赋予对氨基糖苷G418的抗性；潮霉素磷酸转移酶基因，赋予对潮霉素的抗性；ODC基因，赋予对鸟氨酸脱羧酶抑制剂(2-(二氟甲基)-DL-鸟氨酸(DFMO))的抗性；二氢叶酸还原酶基因，赋予对甲氨蝶呤的抗性；嘌呤霉素-N-乙酰转移酶基因，赋予对嘌呤霉素的抗性；ble基因，赋予对博莱霉素的抗性；腺苷脱氨酶基因，赋予对9-β-D-木呋喃糖腺嘌呤的抗性；胞嘧啶脱氨酶基因，允许细胞在N-(膦乙酰基)-L-天冬氨酸的存在下生长；胸苷激酶，允许细胞在氨基蝶呤存在下生长；黄嘌呤-鸟嘌呤磷酸核糖基转移酶基因，允许细胞在存在黄嘌呤和不存在鸟嘌呤的情况下生长；大肠杆菌的trpB基因，允许细胞在存在吲哚而不是色氨酸的情况下生长；大肠杆菌的hisD基因，允许细胞使用组氨醇代替组氨酸。选择基因被掺入到质粒中，该质粒可以另外包含：适于在真核细胞中表达所述基因的启动子(例如，CMV或SV40启动子)、优化的翻译起始位点(例如，遵循所谓的Kozak规则的位点或IRES)、聚腺苷酸化位点(例如SV40聚腺苷酸化)或磷酸甘油酸激酶位点、内含子(例如β-球蛋白基因内含子)。可选择地，可以在同一载体中同时使用报告基因和标记基因的组合。

另一方面，如本领域技术人员所知，载体的选择将取决于随后载体将被导入的宿主细胞。例如，导入所述多核苷酸的载体也可以是酵母人工染色体(YAC)、细菌人工染色体(BAC)或PI衍生的人工染色体(PAC)。YAC、BAC和PAC的特性是本领域技术人员已知的。例如，Giraldo和Montoliu(Giraldo,P.&Montoliu L.,2001Size matters:use of YACs,BACsand PACs in transgenic animals,Transgenic Research 10(2):83-110)已提供了关于所述载体类型的详细信息。本发明的载体可以通过本领域技术人员已知的常规方法获得(Sambrook J.et al.,2000"Molecular cloning,a Laboratory Manual",3rd ed.,ColdSpring Harbor Laboratory Press,N.Y.Vol 1-3)。

本发明的多核苷酸可以作为裸DNA质粒体内导入宿主细胞，也可以通过本领域已知的方法使用载体导入宿主细胞，所述方法包括但不限于转染、电穿孔(例如经皮电穿孔)、显微注射、转导、细胞融合、DEAE葡聚糖、磷酸钙沉淀、使用基因枪、或使用DNA载体转运蛋白。配制裸DNA并将其施用于哺乳动物肌肉组织的方法也是已知的。参见Feigner P等人，US5,580,859和US5,589,466。其他分子也可用于促进核酸的体内转染，例如阳离子寡肽、衍生自DNA结合蛋白的肽、或阳离子聚合物。参见Bazile D等人的WO 1995021931和Byk G等人的WO1996025508。

可用于将多核苷酸导入宿主细胞的另一种众所周知的方法是粒子轰击(又名生物弹道转化(biolistic transformation))。生物弹道转化通常以几种方式之一完成。一种常见的方法涉及在细胞中推入惰性或生物活性颗粒。参见Sanford J等人，US4,945,050、US5,036,006和US5,100,792。

可选择地，可以通过脂质转染将载体进行体内导入。使用阳离子脂质可以促进带负电的核酸的包封，也可以促进与带负电的细胞膜的融合。参见Feigner P，Ringold G，Science 1989；337:387-388。已经描述了用于转移核酸的特别有用的脂质化合物和组合物。参见Feigner P等人的US5,459,127、Behr J等人的WO1995018863和Byk G的WO1996017823。

最后，特别优选的是，可以通过病毒递送系统将载体进行体内导入，所述病毒递送系统包括但不限于腺病毒载体、腺相关病毒(AAV)载体、假型AAV载体、疱疹病毒载体、逆转录病毒载体、慢病毒载体、杆状病毒载体。假型AAV载体是那些在一种AAV血清型的衣壳中包含另一种AAV血清型的基因组的载体；例如，AAV2/8载体包含AAV8衣壳和AAV2基因组(Auricchio et al.(2001)Hum.Mol.Genet.10(26):3075-81)。这样的载体也称为嵌合载体。递送系统的其他实例包括离体递送系统，其包括但不限于DNA转染方法，例如电穿孔、DNA生物弹道技术、脂质介导的转染、压缩DNA介导的转染。

AAV载体的构建可以按照本领域技术人员已知的程序和技术来进行。一些科学和专利出版物中对腺相关病毒载体构建的理论和实践以及在治疗中的应用进行了说明(以下参考书目通过引用并入本文：Flotte TR.Adeno-associated virus-based gene therapyfor inherited disorders.Pediatr Res.2005Dec；58(6):1143-7；Goncalves MA.Adeno-associated virus:from defective virus to effective vector,Virol J.2005May 6；2:43；Surace EM,Auricchio A.Adeno-associated viral vectors for retinal genetransfer.Prog Retin Eye Res.2003Nov；22(6):705-19；Mandel RJ,Manfredsson FP,Foust KD,Rising A,Reimsnider S,Nash K,Burger C.Recombinant adeno-associatedviral vectors as therapeutic agents to treat neurological disorders.MolTher.2006Mar；13(3):463-83)。

含有AAV载体的药物组合物的合适的施用形式包括但不限于可注射溶液或混悬液、眼用洗剂和眼用软膏。因此，另一方面，本发明涉及包含本发明的多核苷酸或载体的宿主细胞。可以通过本领域技术人员已知的常规方法获得细胞(参见例如在上文中引用的Sambrook等人)。

如本文所用，术语“宿主细胞”是指已导入本发明的核酸例如根据本发明的多核苷酸或载体并且能够表达本发明的断裂内含肽N片段的细胞或包含所述断裂内含肽N片段的融合蛋白的细胞。术语“宿主细胞”和“重组宿主细胞”在本文中可互换使用。应当理解，这些术语不仅指特定的对象细胞，而且指这种细胞的后代或潜在后代。由于突变或环境影响，在连续的新一代中可能出现一些修饰，因此，这些后代可能实际上与亲代细胞不相同，但也仍然包含在本文中使用的该术语的范围内。该术语包括任何可通过导入异源DNA进行修饰的可培养的细胞。优选地，宿主细胞是本发明的多核苷酸可以在其中稳定表达、翻译后修饰、定位于合适的亚细胞区室并使其接合合适的转录机制的细胞。选择合适的宿主细胞也会受到检测信号选择的影响。例如，如上所述，报告构建体可在响应转录调节蛋白而激活或抑制基因转录时提供可选择或可筛选的性状；为了实现最佳选择或筛选，将考虑宿主细胞表型。本发明的宿主细胞包括原核细胞和真核细胞。原核生物包括革兰氏阴性或革兰氏阳性生物体，例如大肠杆菌或杆菌(Bacilli)。应当理解，将优选使用原核细胞来增殖本发明的包含转录控制序列的多核苷酸或载体。用于转化的合适的原核宿主细胞包括例如大肠杆菌、枯草芽孢杆菌、鼠伤寒沙门氏菌和假单胞菌属、链霉菌属和葡萄球菌属中的各种其他物种。真核细胞包括但不限于酵母细胞、植物细胞、真菌细胞、昆虫细胞(例如杆状病毒)、哺乳动物细胞和寄生生物(例如锥虫)的细胞。如本文所用，酵母不仅包括严格分类学意义上的酵母，即单细胞生物，还包括丝状真菌的酵母样多细胞真菌。示例性的物种包括乳酸克鲁维酵母(Kluyverei lactis)、粟酒裂殖酵母(Schizosaccharomyces pombe)和玉米黑穗菌(Ustilaqo maydis)，优选的是酿酒酵母。可用于实施本发明的其他酵母是粗糙脉孢菌(Neurospora crassa)、黑曲霉(Aspergillus niger)、构巢曲霉(Aspergillus nidulans)、毕赤酵母(Pichia pastoris)、热带念珠菌(Candida tropicalis)和多形汉逊酵母(Hansenula polymorpha)。哺乳动物宿主细胞培养体系包括已建立的细胞系，例如COS细胞、L细胞、3T3细胞、中国仓鼠卵巢(CHO)细胞、胚胎干细胞，优选BHK、HeK或HeLa细胞。真核细胞优选用于重组基因表达。

在优选的实施方式中，本发明的第二组合物用于治疗，更具体地，用于表1中确定的任何疾病，这取决于组合物中编码的基因类型(疾病和基因之间的相关性在表1中清楚地指出，因此对于本领域技术人员来说识别正确的组合是显而易见的)。

在另一个方面，本发明涉及一种在细胞中表达目标基因的体外或体内方法，下文称为表达目标基因的第一方法，该第一方法包括：

(i)使细胞与以下接触：

(a)本发明的第一多核苷酸或本发明的编码降解决定子的第一多核苷酸，和

(b)本发明的第二多核苷酸或本发明的编码降解决定子的第二多核苷酸，

(ii)使第一多核苷酸和第二多核苷酸表达，以产生第一融合蛋白和第二融合蛋白，以及

(iii)使第一蛋白和第二蛋白接触，使得断裂内含肽N片段与断裂内含肽C片段结合以形成内含肽中间体，并且内含肽中间体反应以将第一目标多肽的C端与第二目标多肽的N端共价连接。

在具体的实施方式中，第一多核苷酸是编码降解决定子的第一多核苷酸，第二多核苷酸是编码降解决定子的第二多核苷酸，其中，优选地，目标蛋白大于25KDa、大于50KDa或大于100KDa，从而在将第一目标多肽的C端与第二目标多肽的N端共价连接后，获得整个蛋白质。

细胞与本发明的任何第一多核苷酸和/或第二多核苷酸的接触可以在体外或体内通过使得目标多核苷酸导入细胞中的任何合适的方法进行，例如转染、电穿孔、显微注射、转导、脂质转染、细胞融合、DEAE葡聚糖、磷酸钙沉淀、使用基因枪

在本发明的优选实施方式中，涉及一种用于在细胞中表达编码目标蛋白的基因的方法，下文称为第一表达目标基因的方法，该方法包括：

(i)使细胞与以下接触：

(a)第一AAV，该第一AAV包含本发明的第一多核苷酸或本发明的编码降解决定子的第一多核苷酸，和

(b)第二AAV，该第二AAV包含本发明的第二个多核苷酸或本发明的编码降解决定子的第二多核苷酸，

(ii)使第一多核苷酸和第二多核苷酸表达，以产生第一融合蛋白和第二融合蛋白，以及

(iii)使第一融合蛋白和第二融合蛋白质接触，使得断裂内含肽N片段与断裂内含肽C片段结合以形成内含肽中间体，并且内含肽中间体反应以将第一目标多肽的C端与第二目标多肽的N端共价连接。

其中优选地，第一多核苷酸是编码降解决定子的第一多核苷酸，第二多核苷酸是编码降解决定子的第二多核苷酸，并且其中，更优选地，目标蛋白大于25KDa、大于50KDa或大于100KDa，从而在将第一目标多肽的C端与第二目标多肽的N端共价连接后，获得整个蛋白质。

从这个意义上说，如实施例中所示，降解决定子的存在将导致起始材料(即本发明的断裂内含肽N和/或C端片段降解决定子)的快速降解。选择的降解决定子具有与蛋白质剪接相容的降解动力学，使得在稳定状态下，相对于没有降解决定子的情况下所观察到的，起始材料的量有所减少。但是，相对于没有降解决定子的情况下所观察到的，剪接产物的水平得以维持或有所增加。

在本发明的表达目标基因的方法中，考虑使细胞与第一多核苷酸和第二多核苷酸同时接触，或以任何顺序依次与第一多核苷酸和第二多核苷酸接触，即可以使细胞首先接触第一多核苷酸，然后再接触第二多核苷酸，或者首先接触第二多核苷酸，然后再接触第一多核苷酸。这同样适用于编码所述多核苷酸的载体。

任何以前定义为宿主细胞的细胞都可以用于这些方法。

***

本发明将通过以下实施例进行描述，这些实施例被认为仅是说明性的，而不是对本发明范围的限制。

实施例

在下表中，我们建立了命名法来表示通过将本文档中已经列出的序列附加在一起而形成的序列。在下表中，通过从N端到C端列出构成它们的以SEQ ID NO识别的序列来表示此类序列。例如，对于SEQ ID NO 106的氨基酸1至1149与SEQ ID NO 27(CfaN)直接连接，而SEQ ID NO 27(CfaN)又与SEQ ID NO103(3FT)直接连接的序列，使用的命名法将是：SEQ IDNO 106(1至1149)-SEQ ID NO 27-SEQ ID NO103。这个具体例子应该被解释为这样的序列：从N端开始，SEQ ID NO 106的氨基酸1至1149，紧随其后的是通过肽键直接连接的对应于SEQ ID NO 27的多肽，其后又紧接着的是也通过肽键连接的对应于SEQ ID NO 103的多肽。因此，序列从N端到C端将具有以下架构：

[SEQ ID NO 106(残基1至1149)]-[SEQ ID NO 27]-[SEQ ID NO 103]

其中括号“[]”中的序列代表该序列中的所有氨基酸，“-”代表连接括号中两个序列的肽键。某个序列的残基x至y，是指从位置x到位置y的所有氨基酸残基，包括位置x和位置y处的氨基酸残基。例如，SEQ ID NO 106(残基1至1149)是指包含来自SEQ ID NO 106的残基1至残基1149(包括残基1和残基1149)的序列。

表5.整个实施例中使用的序列的列表：

寡核苷酸购自Eurofins genomics。合成基因购自GENEWIZ。用于克隆的Pfu超融合聚合酶和所有限制酶均购自Thermofisher Scientific。用于克隆的高效能细胞由XL10-Gold化学感受态大肠杆菌产生。HEK293T细胞购自ATCC。DNA纯化试剂盒购自Qiagen。所有质粒均由Macrogen测序。Luria Bertani(LB)培养基和所有缓冲盐均购自ThermofisherScientific。考马斯亮蓝、MG-132蛋白酶体抑制剂、苯甲磺酰氟、碘乙酰胺、NH4HCO3、DTT、甲酸、胎牛血清和来自大豆的asolectin购自Sigma-Aldrich。乙腈(ACN)购自Carlo-Erba。不含EDTA的完全蛋白酶抑制剂购自Roche。Lipofectamine 2000转染试剂、补充GlutaMAX的高葡萄糖DMEM、补充GlutaMAX的RPMI 1640培养基、RIPA裂解和提取缓冲液、BCA蛋白检测试剂盒、MES-SDS运行缓冲液、预染蛋白梯度物和SDS-PAGE(Bis-tris和Tris-醋酸凝胶)购自Thermofisher Scientific。一抗6xHis标签小鼠单克隆抗体、抗flag标签小鼠单克隆抗体、抗ABCA4兔多克隆抗体和抗微管蛋白兔多克隆抗体购自Invitrogen。山羊抗小鼠IgG(H+L)高度交叉吸附的二抗alexa fluor plus 680和山羊抗兔IgG(H+L)二抗dylight 800 4XPEG购自Invitrogen。十二烷基麦芽糖苷(D310)和胆固醇半琥珀酸酯(CH210)溶液购自Anatrace。胰蛋白酶购自Promega。

电喷雾电离质谱分析(ESI-MS)在与LTQ-Orbitrap Velos(Thermo Scientific)质谱仪耦合的无Acquity液相色谱仪(no Acquity liquid chromatographer)(Waters)上进行。使用LI-COR Odyssey红外成像仪对凝胶和蛋白质印迹进行成像。使用SFX550 Branson超声仪进行细胞裂解。FACS测量在Gallios Beckman Coulter上进行。

购买制备构建体ABCA4-1150-CfaN(SEQ ID NO 1)和ABCA4-1150-CfaC(SEQ ID NO2)的合成基因，并使用KpnI和NotI限制酶将其导入pEGFP-N1表达载体。购买NpuN和NpuC的合成基因，通过无限制酶克隆将其导入ABCA4-1150-CfaN和ABCA4-1150-CfaC，以获得构建体ABCA4-1150-NpuN(SEQ ID NO 8)和ABCA4-1150-NpuC(SEQ ID NO 9)。

通过无限制酶克隆来制备构建体ABCA4-3FT(SEQ ID NO 7)、ABCA4-1140-CfaN(SEQ ID NO 3)、ABCA4-1140-CfaCmut(SEQ ID NO 4)、ABCA4-1188-CfaN(SEQ ID NO 5)、ABCA4-1188-CfaCmut(SEQ ID NO 6)、ABCA4-1179-CfaN(SEQ ID NO 93)和ABCA4-1179-CfaCmut(SEQ ID NO 94)。用Pfu Ultra II HF聚合酶使用反向PCR导入CfaCmut处的突变。通过无限制酶克隆制备ABCA4-1140-NpuN(SEQ ID NO10)、ABCA4-1140-NpuC(SEQ ID NO11)、ABCA4-1188-NpuN(SEQ ID NO 12)和ABCA4-1188-NpuC(SEQ ID NO 13)。

通过无限制酶克隆制备构建体EGFP-71-CfaN(SEQ ID NO 14)、EGFP-71-CfaC(SEQID NO 15)、EGFP(SEQ ID NO 16)、EGFP-71-NpuN(SEQ ID NO 17)和EGFP-71-NpuC(SEQ IDNO 18)。

购买SopE的合成基因，并通过无限制酶克隆将其导入ABCA4-1150-CfaN、ABCA4-1150-CfaC、ABCA4-1140-CfaN、ABCA4-1140-CfaC、EGFP-71-CfaN和EGFP-71-CfaC，以获得构建体ABCA4-1150-CfaN-SopE(SEQ ID NO 19)、ABCA4-1150-CfaC-SopE(SEQ ID NO 20)、ABCA4-1140-CfaN-SopE(SEQ ID NO 21)、ABCA4-1140-CfaC-SopE(SEQ ID NO 22)、EGFP-71-CfaN-SopE(SEQ ID NO 23)和EGFP-71-CfaC-SopE(SEQ ID NO 24)。通过重叠延伸PCR制备DD1基因，并通过无限制酶克隆将其导入EGFP-71-CfaN和EGFP-71-CfaC，以得到构建体EGFP-71-CfaN-DD1(SEQ ID NO25)和EGFP-71-CfaC-DD1(SEQ ID NO 26)。通过测序确认所有重组质粒的身份，表1中报告了相应的蛋白质序列。通过将包括断裂蛋白基因、断裂内含肽和降解决定子的不同元件克隆到表达质粒中类似地产生了具有降解决定子的构建体。

使用标准三质粒转染(triple transfection)策略产生编码本发明的N片段(编码具有CfaN内含肽的EGFP或ABCA4片段)的AAV。具有野生型AAV2 ITR的rAAV8载体是通过聚乙烯亚胺(PEI)介导在HEK293细胞中共转染质粒pAAV-Rep2-Cap8、pHelper和AAV-转基因-质粒产生的。

转染后48小时收获细胞和培养基。在用0.1％Triton处理后，病毒从细胞中释放出来，并且通过PEG沉淀从培养基中获得AAV颗粒。根据Zolotukhin及其同事的方法[Zolotukhin S,Byrne BJ,Mason E,Zolotukhin I,Potter M,Chestnut K,Summerford C,Samulski,RJ,Mucyczka N.(1999)Recombinant adeno-associated virus purificationusing novel methods improves infectious titer and yield.Gene Ther；6(6):973-85.]通过碘克沙醇梯度纯化粗裂解液并去除空衣壳。

使用Vivaspin 20离心浓缩器100.000MWCO(Sartorius，目录号VS0641)将纯化的批次浓缩并配制至1×10

浓缩后，使用Acrodisc(R)注射器过滤器(Acrodisc PP，PES，0.2μM 1cm2)过滤病毒批次，并储存在-80℃。使用ITR特异性PCR引物通过qPCR来确定病毒滴度，病毒滴度以每毫升基因组拷贝数表示。此外，衣壳滴度是使用AAV8 ELISA(Progen)根据制造商的说明获得的。

使用Pierce Silver染色试剂盒(Cat#24612)在10％ SDS PAGE上分离存在于最终产物中对应于1×10

HEK293T细胞在37℃和6％CO2气氛下在含有10％FBS和抗生素的DMEM中维持。以6孔板的形式，使用Lipofectamine 2000和1.25μg各质粒在汇合度约80％时共转染细胞。对于使用编码全长基因的质粒的实验，当使用两个内含肽质粒时，将乱序质粒(scrambleplasmid)与全长质粒共转染以实现相同量的转染的DNA。转染后48小时收获细胞，并通过蛋白质印迹分析EGFP。

以6孔板的形式，使用Lipofectamine 2000和1.25μg各质粒在汇合度约80％时共转染细胞。转染后48小时收获用内含肽-降解决定子质粒转染的细胞并通过蛋白质印迹分析。对于时程实验，在转染后24小时和48小时后收获细胞并通过蛋白质印迹分析。

使用MG-132抑制剂进行蛋白酶体抑制剂实验。共转染细胞并在转染后24小时加入DMSO或MG-132(溶解在DMSO中)至终浓度为50μM。在30分钟、3小时、6小时和24小时后收获细胞并通过蛋白质印迹分析。

将EGFP转染细胞(HEK293和WERI-RB1细胞)在补充有蛋白酶抑制剂和1mM苯甲基磺酰的RIPA缓冲液中裂解。裂解后，通过BCA蛋白质测定试剂盒对样本进行定量。将含有10μg总蛋白的样本在1XLaemmli样本缓冲液中于95℃变性10分钟。将裂解物通过12％ Bis-trisSDS-PAGE凝胶在165V下分离40分钟。用于免疫印迹的抗体是抗6xHis标签抗体，以检测EGFP和作为上样对照的抗β-微管蛋白。使用LI-COR Odyssey红外成像仪对通过蛋白质印迹检测到的EGFP条带进行定量。

将ABCA4转染细胞(HEK293)在十二烷基麦芽糖苷(D310)和胆固醇半琥珀酸酯(CH210)的PBS溶液(1:1)(补充有蛋白酶抑制剂和1mM苯甲基磺酰基)中裂解。裂解后，ABCA4样本通过BCA蛋白检测试剂盒进行定量。含有25μg总蛋白的样本在含有2.5mg/mlasolectin的1X Laemmli样本缓冲液中于37℃变性15分钟。将裂解物通过3％-8％Tris-乙酸盐SDS-PAGE凝胶在150V下分离1.5小时。用于免疫印迹的抗体是抗flag标签或抗ABCA4抗体，以检测ABCA4蛋白和作为上样对照的抗β-微管蛋白。使用LI-COR Odyssey红外成像仪对通过蛋白质印迹检测到的ABCA4带进行定量。

HEK293T细胞用2μg各EGFP-内含肽质粒共转染，作为阴性对照的HEK293T细胞生长但不进行转染。对于使用编码全长基因的质粒的实验，当使用两个内含肽质粒时，将乱序质粒与全长质粒共转染以实现相同量的转染的DNA。转染后48小时后通过流式细胞术分析细胞。转染和未转染的细胞分别重悬于FACS分析缓冲液(PBS，2％FSA，2mM EDTA)中。通过使用Kaluza流式细胞术分析软件在Beckman Coulter Gallios流式细胞仪中对不同的转染细胞和未转染细胞进行比较来评估EGFP+细胞的百分比。使用Dapi染色来检测死细胞的数量。

用碳酸氢铵(50mM NH4HCO3)和乙腈(ACN)洗涤凝胶条带。样本在60℃下用20mMDTT还原60分钟，然后在25℃下用55mM碘乙酰胺避光进行烷基化30分钟。之后，将样本在37℃下用胰蛋白酶(序列级改良胰蛋白酶)双重消化2小时并过夜。最后，用5％甲酸(FA)的50％ACN溶液、和100％ACN从凝胶基质中提取得到的肽混合物，并在SpeedVac真空系统中干燥。根据制造商的方案，使用C18吸头(PolyLC Inc.)对所得的肽混合物进行清洗。最后，将清洗后的肽溶液干燥。将胰蛋白酶消化混合物重悬于1％FA溶液中，对于每个样本，注入等分试样进行色谱分离。将肽捕获在Symmetry C18捕集柱(5μm180μm×20mm；Waters)上，并使用C18反相毛细管柱(ACQUITY UPLC M-Class Peptide BEH柱；

用4周龄的129Sv品系野生型动物进行实验。用测试样本对小鼠进行视网膜下注射。

在瞳孔扩张后使用5ml Hamilton注射器上的33G针头进行视网膜下注射。使用与数字成像系统连接的佳能UVI视网膜相机实现了注射后4周的EGFP表达眼底评估。使用Spectralis成像系统(Heidelberg Engineering Inc.)通过SD-OCT(光学相干断层扫描)来评估视网膜结构和外核层(ONL)的定量。

成像后，对动物实施安乐死，摘除眼睛以通过免疫组织化学和蛋白质印迹进行进一步分析。进行了另外的实验，其中将载体注射到6周C57BL/6小鼠的内耳中。

最近，基于共有设计(consensus design)而工程改造了若干新的内含肽((Stevens et al.,2016；Stevens,Sekar,Gramespacher,Cowburn,&Muir,2018))并证明其具有优于天然存在的内含肽的特性。这些内含肽中一种被称为Cfa的内含肽是由DnaE内含肽的比对通过共有设计产生的，并显示出比DnaE家族中一些性能最好的内含肽(如Npu)更优越的特性。据报道，Cfa具有更快的动力学、更高的表达水平和对极端条件(例如高温和变性剂浓度)的高耐受性。有趣的是，具有90％或更高程度的同源性的Cfa变体显示出相似的特性。将相同的共有设计策略应用于TerL-AceL内含肽家族，产生了称为Cat的共有序列，相对于TerL-AceL家族的其他成员，Cat也显示出改进的特性。

为了证明共有内含肽通过蛋白质反式剪接为蛋白质重构提供了用于基因治疗应用的益处，进行了直接比较。通过用纯化的质粒转染细胞进行比较。

最初的实验是使用EGFP作为报告基因进行的。简而言之，EGFP在位置71处断裂，N端片段(残基1-70，EGFPN)与N内含肽重组融合，而C端片段(残基71-239，EGFPC)与C内含肽重组融合。生成了以下四种构建体：EGFPN-CfaN、EGFPN-NpuN、CfaC-EGFPC和NpuC-EGFPC，从而比较Cfa共有序列与Npu的重构效率。用等摩尔量的编码N端片段和C端片段的质粒共转染培养的HEK293细胞，并通过荧光显微镜、流式细胞术分析和蛋白质印迹监测剪接效率。

我们的结果(参见图2、图3和图4)表明，与Npu内含肽相比，共有Cfa内含肽使EGFP重构增加至2.5倍，这表明通过共有设计获得的内含肽在共转染时提供了更高产量的目标蛋白。需要指出的是，尽管已知Cfa在体外以比Npu更快的速度断裂，它的N端片段在单独转化或转染到细胞中时表达更好，但是当两个片段共转染到同一个细胞中时，这是否会带来益处尚未被证明。通过这些实验，我们证明，当IntN和IntC片段在同一细胞中表达时，与先前已知的超快速断裂内含肽(例如Npu)相比，使用Cfa在目标蛋白的重构产量方面提供了明显的益处。编码EGFP的N片段和C片段以及N内含肽和C内含肽的基因也被掺入到重组AAV中用于其体内递送。结果表明，Cfa内含肽介导的蛋白质剪接导致EGFP在包括视网膜和内耳在内的几个器官中重构(图23)。

为了证实在其他蛋白质中观察到Cfa共有序列的这种积极特征，我们使用ABCA4蛋白质重复了实验。ABCA4是一种大蛋白，其在Stargardt病中发生突变，并且ABCA4重构已被提议作为治疗该疾病的可行策略。目前正在探索几种基于AAV基因治疗的方法对ABCA4进行重构。由于其大小较大，ABACA4不能封装到单个AAV中，因此已经提出了不同的策略以便能够将ABCA4以两个片段进行递送，从而在靶细胞内重构。为了确认工程改造的共有序列(如Cfa)提供了优于天然存在的内含肽(如Npu)的益处，我们在位置1150处使ABCA4断裂，并将N内含肽和C内含肽分别克隆到蛋白的N端片段和C端片段。

具有Cfa内含肽或Npu内含肽的ABCA4N(1至1149)-IntN和IntC-ABCA4

正如对EGFP观察到的那样，我们观察到Cfa的重构产量高于Npu(见图5)。

最近还报道了一种Cfa变体，它在CfaC片段中包含特定突变，称为CfaCmut((Stevens et al.,2017))，它可以增加Cfa的益处。特别是CfaCmut变体已被证明能够有效地剪接蛋白质片段，即便C外显肽的+2位置不包含天然Phe残基。如上定义的+2位置是指位置+2(相对于IntC的最后一个氨基酸)。当ABCA4在位置1150处断裂时，在C外显肽+2位置为Phe，其对于Npu和Cfa是最佳残基。

尽管可以通过在位置1150位处使ABCA4断裂来重构ABCA4，但这可能不是用于治疗目的的最佳位置。为了测试我们通过在1150以外的不同位点使ABCA4断裂来重构ABCA4的能力，我们使用了CfaCmut，以便不限制于在+2位置(相对于内含肽)处呈现Phe的位置。

克隆了若干ABCA4构建体，其中ABCA4蛋白在断裂位点的位置+2处不包含Phe的位点断裂。测试的另外两个位点是位置1140，其对应于+2位点处为Ser，以及位置1188，其对应于+2位点处为Pro。如上所示进行克隆并测试构建体。具体而言，使用ABCA4(1至1139)-IntN、IntC-ABCA4(1140至2273)构建体来评估ABCA4在位置1140处的剪接。使用ABCA4(1至1187)-IntN和IntC-ABCA4(1188至2273)来评估在位置1188处使ABCA4断裂。根据ABCA4的拓扑结构并考虑折叠结构域的存在来选择位点。在ABCA4细胞内侧明确定义的折叠结构域之外选择位点。对不同的位点进行了测试，对所有这些位点都观察到Cfa和Cfa变体提供比Npu更高的重构产量，并且效率也因断裂位点而异(图5、6和7)。我们还证明了该方法允许ABCA4在位置1179和1177处断裂重构。克隆了与上面1140和1188所定义的类似的构建体，并用于测试位置1179和1177处的剪接效率，位置1179的结果显示在图21中。

我们还证明了其他有效的内含肽(如gp41)可用于由断裂片段重构ABCA4。设计了在位置1185和1095处断裂的ABCA4构建体，克隆了与N端内含肽和C端内含肽的相应融合物，并如上所述分析了它们的剪接活性。通过蛋白质印迹对断裂位置1096处的PTS反应和gp41内含肽的分析如图22所示。

下表7示出了当蛋白质在不同位点(1177、1179、1096和1185)断裂时获得的ABCA4片段的序列以及相应的ABCA4-内含肽N或内含肽C-ABCA4融合体。

除了重构产量之外，在基因治疗中使用断裂内含肽的另一个主要限制是存在未反应的起始材料和/或内含肽。为了防止不需要的起始材料的积累并消除切下的内含肽片段，我们向构建体添加了降解决定子，一般架构如图8所示。

我们已经确定了几种可以与内含肽结合使用的降解决定子，以开发可以转化出由其他大基因突变引起的疾病的ABCA4的基因治疗方法以及重构蛋白质来治疗这些疾病，例如重构CRISPR/Cas9系统。

为了识别合适的降解决定子-内含肽组合，克隆了将选定的降解决定子包括在内的EGFP-Int构建体。在N内含肽的C端和C内含肽的N端克隆了降解决定子。为了检测目的，将His6标签包含在两个元件之间作为接头。为了证明原理，使用了SopE去稳定结构域(1-100)，以及由肽序列DD1组成的降解决定子(参见表3)

结果表明，包含降解决定子确实去除了任何可检测量的起始材料和内含肽(参见图9、图14)。重要且出乎意料的是，事实证明，同时包含两种降解决定子可以去除任何未反应的起始材料，而不会损失蛋白质剪接反应的产率。

有趣的是，当降解决定子策略应用于大基因ABCA4时，我们观察到类似的结果，而且剪接ABCA4产物的水平也有所增加，这表明共有内含肽和降解决定子之间存在出乎意料的协同效应(见图10)。

还研究了蛋白体抑制剂对这些结果的影响(图11)。我们发现蛋白体抑制剂降低了降解决定子的作用，从而证实了SopE降解是蛋白酶体介导的。有趣的是，我们观察到将内含肽与降解决定子相结合，我们不仅能够消除起始材料的存在，而且能够维持甚至提高ABCA4的重构水平(图10)。

我们还进行了实验来研究将共有内含肽Cfa与降解决定子相结合的效果，并将其与不存在降解决定子或使用其他内含肽进行比较。有趣的是，我们观察到(图12)将Cfa与降解决定子一起使用非常显著地降低了起始材料的水平，并且还增加了产物的量。我们还确认了在ABCA4的位置1140(图13和图21)和1179(图21)也观察到了这种效果。重要的是，我们表明，将Cfa(和Cfamut)内含肽与几种不同的降解决定子(这些降解决定子具有某些共同的特征)结合所观察到的效果得以维持(参见实施例3)。

基于ABCA4获得的结果，可以应用该方法的其他疾病、基因和蛋白质包括表1中所示的那些。

在HEK293细胞中转染三段式质粒：

以6孔板的形式，使用Lipofectamine 2000和1.25μg各质粒在汇合度约80％时共转染细胞。转染后48小时收获用三段式质粒转染的细胞并通过蛋白质印迹分析。作为对照，用两种质粒(POIN-CfaN+CfaC-POIM-CatN、CfaC-POIM-CatN+CatC-POIC和POIN-CfaN+CatC-POIC)共转染细胞。

将三段式转染细胞(HEK293)在补充有蛋白酶抑制剂和1mM苯甲基磺酰的RIPA缓冲液中裂解。裂解后，通过BCA蛋白质测定试剂盒对样本进行定量。将含有10μg总蛋白的样本在1X Laemmli样本缓冲液中于95℃变性10分钟。将裂解物通过12％Bis-tris SDS-PAGE凝胶在165V下分离40分钟。用于免疫印迹的抗体是抗flag标签抗体，以检测POI和作为上样对照的抗β-微管蛋白。

我们表明，将超快共有内含肽与降解决定子组合是重构大蛋白的可行策略，同时降低了起始材料和切下的内含肽的水平。该策略可用于基因治疗，以在基因替换策略中重构大蛋白，以及在与合适的递送载体(例如本文公开的递送载体)组合时在体内产生治疗剂。然而，对于更大的蛋白质，这种策略可能还不够。例如，对于编码区大于7kb-8kb的蛋白质，将其编码基因断裂成两个片段不足以产生可封装在AAV载体中的片段。对于这样的蛋白质，编码基因可能需要断裂成三段。为了使用内含肽从三个单独的片段组装蛋白质，需要正交内含肽对。为实现最大重构产量，需要高效的正交内含肽。我们决定将Cfa与Cat内含肽结合使用。这两个内含肽是通过共有设计获得的，并具有一些共同特征，包括高表达产量、热稳定性、对离液剂的耐受性和快速的剪接动力学。我们按照图16中描述的结构设计了一系列构建体：POIN-CfaN、CfaC-POIM-CatN和CatC-POIC。可选择地，可以交换Cfa和Cat内含子的位置，以生成具有以下架构的构建体：POIN-CatN、CatC-POIM-CfaN和CfaC-POIC。

我们测试了它们的正交性，证明了两个内含肽确实是正交的，它们不相互反应，即Cfa或Cat内含肽的N片段仅与其各自的同源对反应。当三个片段以等摩尔量共转染时，检测到的主要产物是由三个片段组装成全长期望产物而产生的产物。重要的是，我们只检测到低水平的未反应材料，我们没有检测到任何中间产物，即仅由两个片段反应产生的产物。该结果表明使用高活性正交共有内含肽对是由三个单独的片段组装大蛋白的合适策略。

为了实践本发明，测试了若干个降解决定子。根据它们的大小从文献中选择降解决定子，对于AAV基因治疗应用，小于75个氨基酸的降解决定子将是优选的，以使所得转基因的大小最小化。在某些情况下，降解决定子是通过组合已知的N端降解决定子或C端降解决定子来设计的。

在N片段的C端和C片段的N端克隆了降解决定子，这两个片段都旨在重构全长EGFP蛋白，作为可溶性细胞溶质蛋白的代表性例子。在图9中可以观察到结果的总结，其中可以看出包含一个或两个降解决定子(每个EGFP片段上一个)消除了不需要的起始材料片段，而不会对剪接产量产生负面影响。使用不同的降解决定子观察到类似的结果(图14)，说明了该方法的通用性。

在N片段的C端和C片段的N端克隆了降解决定子，这两个片段旨在在不同的断裂位置(作为代表性示例，包括1140、1150和1179)重构全长ABCA4蛋白。下面的表6示出了所有测试的降解决定子，包括大小、序列、它们的来源以及负责它们泛素化和最终降解的连接酶。

对降解决定子进行测试以确认它们是否能够诱导起始材料和切离的内含肽的降解，并确认它们对蛋白质反式剪接反应产量的影响，即它们重构全长ABCA4蛋白的能力。用含有上述不同降解决定子的N端构建体和C端构建体共转染细胞。裂解细胞并通过蛋白质印迹分析以检测具有和不具有降解决定子的PTS产物的存在。在图18中，示出了在N片段和C片段上使用相同降解决定子的结果。左图(图18C)上的凝胶显示了每个反应的PTS产物(ABCA4)以及起始材料(ABCAN-IntN-DD和DD-IntC-ABCAC)。PTS对应于没有使用降解决定子时的结果，其他泳道中的标签指示在每种情况下使用了哪种降解决定子。右图(图18C)提供了每种情况下PTS产物量的量化。可以看出，令人惊讶的是，与不具有降解决定子的构建体相比，一些降解决定子提供更高的产量。有趣的是，所有的降解决定子都有效，并且使用所有的降解决定子都可以重构在不具有降解决定子的情况下获得的产物的至少25％。一些降解决定子允许重构在不具有降解决定子的情况下获得的水平的超过50％，最后，有一些降解决定子产生100％(或更高)的重构。具体而言，产生100％(或更高)重构的降解决定子如下：DD1、V12、M4、L2、L9、DD3和SopE100。

还研究了降解决定子对切下的内含肽的影响(图19)。使用高百分比丙烯酰胺凝胶检测内含肽，通过WB(蛋白质印迹)分析具有不同降解决定子的PTS反应。可以看出，与其他降解决定子(例如SpoE和PEST)相比，几种降解决定子减少了切下的内含肽的量，而其他降解决定子(如L2(SEQ ID NO 52)、L9(SEQ ID NO 54)、M2(SEQ ID NO 47)、M4(SEQ ID NO35)或DD3(SEQ ID NO 45))则完全消除了内含肽条带。

此外，我们还进行了实验以研究在N端位置或C端位置使用不同的降解决定子的影响。例如，在图20中，我们示出了蛋白质印迹分析，其中N片段处的降解决定子是固定的，而C片段处的降解决定子是变化的。图20中所示的结果表明，将一些优选的降解决定子组合可以获得进一步改进的结果，从而在不降低所需剪接产物的产量的情况下实现了起始材料的进一步减少。这些组合可能对某些应用，特别是当完全消除任何起始材料至关重要时是有用的。

根据获得的结果，我们可以将降解决定子分为三类：

·使得能够以没有降解决定子时获得的产量的至少25％重构目标蛋白的降解决定子。

·使得能够以没有降解决定子时获得的产量的约50％以上的产量重构目标蛋白的降解决定子。

·使得能够以等于或优于没有降解决定子时获得的产量的产量重构目标蛋白的降解决定子。

当使用CfaN和CfaC内含肽或CfaCmut内含肽的组合时，表明ABCA4中的几个位点适合于在此描述的本发明的使用。具体来说，是核苷酸结合结构域1(NBD1)的C端与第七跨膜结构域(TMD)之间的位点，包括位置1140、1150、1177、1179和1188。ABCA4重构产量在位置1140、1150和1179处是最佳的，虽然位置1188的工作效率非常低，但仍然导致ABCA4重构水平高于使用非共有Npu内含肽获得的重构水平。

CfaN和CfaCmut内含肽的组合提供了增加的混杂性。事实上，我们在这里首次展示了这种增加的混杂性是在跨膜蛋白(如ABCA4)中观察到的，其中在不同剪接位点使用CfaN-CfaCmutant对获得了高重构率。此外，当使用这些内含肽(CfaN和CfaCmut内含肽)时，这种剪接反应显示出非常有效，因为观察到的每种起始材料不到10％(如图12中所述估计)。这是根据先前报道的使用Npu内含肽与ABCA4的结果(Auricchio et al.2019)所无法预期的，因为其中很大一部分起始材料仍然没有反应，这表明蛋白质反式剪接对于这种膜蛋白来说本来就更具挑战性。

根据此处报告的结果，我们得出结论，当使用共有内含肽对CfaN和CfaCmutant时，具有以下特征的任何位点都将有效工作：(1)酶促结构域或跨膜结构域之外的位点；(2)诸如断裂位点的+1位置(如上所述)的位点应该对应于亲核残基(如Cys或Ser)，并且+2位置可以是除Pro外的任何位点。

1、一种组合物，该组合物包括第一多核苷酸和第二多核苷酸，所述第一多核苷酸编码包含通过肽键、可选地通过肽接头与要重构的蛋白质的N端片段直接连接的断裂内含肽N片段的多核苷酸，所述第二多核苷酸编码包含通过肽键、可选地通过肽接头与要重构的蛋白质的C端片段直接连接的断裂内含肽C片段的多核苷酸；

其中所述组合物的两种组分可以一起包装在单一制剂中或分别包装在不同制剂中；

其中所述第一多核苷酸和所述第二多核苷酸分别编码所述要重构的蛋白质的所述N端片段和所述C端片段，使得当两个片段组合时，所述蛋白质的所述N端片段与所述蛋白质的所述C端片段连接，从而产生整个蛋白质；

其中各多核苷酸必须编码断裂内含肽，使得一旦翻译成蛋白质，所述N端序列和所述C端序列会变成可非共价重新缔合或重构为对反式剪接反应起作用的内含肽的单独的片段；以及其中

所述要重构的蛋白质大于25KDa。

2、根据条款2所述的组合物，其中，所述第一多核苷酸编码通过肽键与降解决定子直接连接的断裂内含肽N片段，其中所述降解决定子在所述内含肽N片段和所述降解决定子之间有或没有接头的情况下通过内含肽的C端与所述内含肽N片段连接，并且其中所述断裂内含肽N片段的N端通过肽键与所述要重构的蛋白质的N端片段直接连接；以及其中，所述第二多核苷酸编码通过肽键与降解决定子直接连接的断裂内含肽C片段，其中所述降解决定子在内含肽C片段和降解决定子之间有或没有接头的情况下通过内含肽的N端与所述内含肽C片段连接，并且其中所述断裂内含肽C片段的C端通过肽键与所述要重构的蛋白质的C端片段直接连接。

3、根据条款1或2中任一项所述的组合物，其中，所述第一多核苷酸编码SEQ ID NO27的CfaN内含肽或其任何变体，并且所述第二多核苷酸编码SEQ ID NO 28的CfaC内含肽或其任何变体，其中变体被理解为与这些序列中的任何序列具有至少90％序列同一性的SEQID NO：27或SEQ ID NO 28的断裂内含肽N片段或C片段，如CfaCmut(SEQ ID NO 29)。

4、根据条款2或3中任一项所述的组合物，其中，降解决定子选自由SEQ ID NO 40-59、34、35和37组成的清单。

5、根据条款2所述的组合物，其中，所述断裂内含肽如权利要求3所定义，并且其中所述降解决定子选自由SEQ ID NO 42、43、49、50、51、54和34组成的清单。

6、根据条款1至5中任一项所述的组合物，其中，两种多核苷酸都包含在使得所述多核苷酸在合适的宿主细胞中增殖的载体中。

7、根据条款6所述的组合物，其中，所述载体是腺相关病毒(AAV)。

8、根据条款7所述的组合物，其中，所述载体是血清型1、2、3、4、5、6、7、8或9的AAV。

9、根据条款1至8中任一项所述的组合物，其中，编码整个蛋白质的基因选自由表1中列出的任何蛋白质组成的清单。

10、用于治疗的根据条款1至9中任一项所定义的组合物。

11、一种在细胞中表达目标基因的方法，包括：

(i)使细胞与以下接触：

(a)条款1中定义的第一多核苷酸，和

(b)条款1中定义的第二多核苷酸，

(ii)使第一多核苷酸和第二多核苷酸表达，以产生第一融合蛋白和第二融合蛋白，以及

(iii)使第一蛋白和第二蛋白接触，使得断裂内含肽N片段与断裂内含肽C片段结合以形成内含肽中间体，并且内含肽中间体反应以将第一目标多肽的C端与第二目标多肽的N端共价连接。

12、一种在细胞中表达目标基因的方法，包括：

(i)使细胞与以下接触：

(a)条款2中定义的第一多核苷酸，和

(b)条款2中定义的第二多核苷酸，

(ii)使第一多核苷酸和第二多核苷酸表达，以产生第一融合蛋白和第二融合蛋白，以及

(iii)使第一蛋白和第二蛋白接触，使得断裂内含肽N片段与断裂内含肽C片段结合以形成内含肽中间体，并且内含肽中间体反应以将第一目标多肽的C端与第二目标多肽的N端共价连接。

13、根据条款11或12中任一项所述的方法，其中，所述第一多核苷酸编码SEQ IDNO 27的CfaN内含肽或其任何变体，并且所述第二多核苷酸编码SEQ ID NO 28的CfaC内含肽或其任何变体，其中变体被理解为与这些序列中的任何序列具有至少90％序列同一性的SEQ ID NO：27或SEQ ID NO 28的断裂内含肽N片段或C片段，如CfaCmut(SEQ ID NO 29)。

14、根据条款11或12中任一项所述的方法，其中，所述断裂内含肽如权利要求3所定义，并且其中所述降解决定子选自由SEQ ID NO 42、43、49、50、51、54和34组成的清单。

15、根据条款11至14中任一项所述的方法，其中，两种多核苷酸都包含在腺相关病毒(AAV)中。

序列表

<110> 移接生物有限公司

<120> 断裂内含子及其用途

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<140> EPPCT/2021/058016

<141> 2020-03-26

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Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

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Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

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Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

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Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

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Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

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Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

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Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

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Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

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Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

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Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

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Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

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His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

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Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

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Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

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Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

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Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

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His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

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Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

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Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

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Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

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Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

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Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

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Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

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Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

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Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

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Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

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Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

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Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

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Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

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Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

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Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

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Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

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Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

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Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

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Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

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Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

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Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

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Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

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Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

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His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

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Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

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Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

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Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

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Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

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Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

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Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

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Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

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Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

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Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

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Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

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Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

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Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

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Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

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Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

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Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

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Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

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Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

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Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

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Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

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Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

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Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

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Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

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Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

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Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Leu Ser

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Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Phe Leu Pro Ile Gly

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Lys Ile Val Glu Glu Arg Ile Glu Cys Thr Val Tyr Thr Val Asp Lys

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Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala Gln Trp His Asn Arg Gly

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Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser Ile Ile Arg

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Ala Thr Lys Asp His Lys Phe Met Thr Thr Asp Gly Gln Met Leu Pro

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Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp Leu Lys Gln Val Asp Gly

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Leu Pro Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile

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Asp Tyr Lys Asp Asp Asp Asp Lys

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Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

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Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu Lys Asn Gly Leu

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Val Ala Ser Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg

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Lys Met Lys Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys

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Ser Cys Ser Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp

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Asp Leu Thr Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met

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Asp Val Val Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile

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Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg

115 120 125

Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu

130 135 140

Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe

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Leu Lys Val Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly

165 170 175

Ala Gln Gln Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly

180 185 190

Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser

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Pro Gly Ala Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro

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Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln

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His Val Gln Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser

245 250 255

His Lys Asp Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe

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Leu Ala Leu Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro

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Ala Leu Thr Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe

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Ser Met Asp Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val

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Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp

325 330 335

Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser

340 345 350

Val Ser Pro Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln

355 360 365

Val Asn Pro Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr

370 375 380

Met Leu Pro Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln

385 390 395 400

Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn

405 410 415

Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser

420 425 430

Leu Lys Ser Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser

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Ile Gly Gly Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val

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Gly Phe Leu Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro

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Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu

485 490 495

Glu Thr Glu Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His

500 505 510

Ala Leu Val Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala

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Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile

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Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val

545 550 555 560

Leu Thr Thr Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser

565 570 575

Met Ser Phe Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg

580 585 590

Val Asn Lys Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr

595 600 605

Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val

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Ser Ala Gly Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala

625 630 635 640

Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu

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Tyr Gly Trp Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe

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Asp Val Pro Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe

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Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu

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Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu

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Ile Val Phe Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala

725 730 735

Leu Ser Gln Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His

740 745 750

Ser Ala Asn Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala

755 760 765

Met Val Val Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln

770 775 780

Arg His Phe Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro

785 790 795 800

Ile Val Asp Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile

805 810 815

Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys

820 825 830

Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly

835 840 845

Val Arg Pro Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly

850 855 860

Lys Thr Thr Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser

865 870 875 880

Gly Asp Ala Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu

885 890 895

Val His Gln Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu

900 905 910

Leu Leu Thr Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly

915 920 925

Val Pro Ala Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser

930 935 940

Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly

945 950 955 960

Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro

965 970 975

Pro Leu Val Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala

980 985 990

Arg Arg Met Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg

995 1000 1005

Ala Val Val Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys

1010 1015 1020

Thr Arg Leu Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr

1025 1030 1035 1040

Ile Gln His Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met

1045 1050 1055

Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val

1060 1065 1070

Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu Arg

1075 1080 1085

His Tyr Asn Met Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala Arg

1090 1095 1100

Ile Phe Gln Leu Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu Glu

1105 1110 1115 1120

Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe Ala

1125 1130 1135

Lys Gln Gln Thr Glu Ser His Asp Leu Pro Leu His Pro Arg Ala Ala

1140 1145 1150

Gly Ala Ser Arg Gln Ala Gln Asp Asp Tyr Lys Asp His Asp Gly Asp

1155 1160 1165

Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys

1170 1175 1180

<210> 3

<211> 1262

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaN-3FT

<400> 3

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr

1140 1145 1150

Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr

1155 1160 1165

Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala

1170 1175 1180

Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1185 1190 1195 1200

Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr

1205 1210 1215

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp

1220 1225 1230

Leu Lys Gln Val Asp Gly Leu Pro Asp Tyr Lys Asp His Asp Gly Asp

1235 1240 1245

Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys

1250 1255 1260

<210> 4

<211> 1192

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaCmut-3FT

<400> 4

Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Gly Glu Pro His Asn Phe Leu Leu Lys Asn Gly Leu

20 25 30

Val Ala Ser Asn Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe

35 40 45

Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln

50 55 60

Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly

65 70 75 80

Phe Ser Thr Thr Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln

85 90 95

Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val Leu His His

100 105 110

Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe

115 120 125

Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe

130 135 140

Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly

145 150 155 160

Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp

165 170 175

Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu

180 185 190

Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly

195 200 205

Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala

210 215 220

His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln

225 230 235 240

Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu

245 250 255

Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala

260 265 270

Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser

275 280 285

Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro

290 295 300

Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly

305 310 315 320

Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly

325 330 335

Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys

340 345 350

Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr

355 360 365

Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser

370 375 380

Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro

385 390 395 400

Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr

405 410 415

Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val

420 425 430

Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp

435 440 445

Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro

450 455 460

Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu

465 470 475 480

Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser

485 490 495

Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile

500 505 510

Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu

515 520 525

Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg

530 535 540

Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu

545 550 555 560

Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp

565 570 575

Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala

580 585 590

Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His

595 600 605

Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn

610 615 620

Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val

625 630 635 640

Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn

645 650 655

Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile

660 665 670

Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala

675 680 685

Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser

690 695 700

Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu

705 710 715 720

Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe

725 730 735

Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr

740 745 750

Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His

755 760 765

Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val

770 775 780

Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser

785 790 795 800

Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp

805 810 815

Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr

820 825 830

Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser

835 840 845

Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys

850 855 860

Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys

865 870 875 880

Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala

885 890 895

Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly

900 905 910

Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu

915 920 925

His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile

930 935 940

Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr

945 950 955 960

Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu

965 970 975

Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp

980 985 990

Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn

995 1000 1005

Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser

1010 1015 1020

His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met

1025 1030 1035 1040

Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser

1045 1050 1055

Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys

1060 1065 1070

Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly

1075 1080 1085

Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln

1090 1095 1100

Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu

1105 1110 1115 1120

Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr

1125 1130 1135

Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser

1140 1145 1150

His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala

1155 1160 1165

Gln Asp Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile

1170 1175 1180

Asp Tyr Lys Asp Asp Asp Asp Lys

1185 1190

<210> 5

<211> 1310

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1188-CfaN-3FT

<400> 5

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly

1140 1145 1150

Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser

1155 1160 1165

Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe

1170 1175 1180

Ser Thr Thr Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr

1185 1190 1195 1200

Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr

1205 1210 1215

Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala

1220 1225 1230

Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1235 1240 1245

Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr

1250 1255 1260

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp

1265 1270 1275 1280

Leu Lys Gln Val Asp Gly Leu Pro Asp Tyr Lys Asp His Asp Gly Asp

1285 1290 1295

Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys

1300 1305 1310

<210> 6

<211> 1144

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1188-Cfacmut-3FT

<400> 6

Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Gly Glu Pro His Asn Phe Leu Leu Lys Asn Gly Leu

20 25 30

Val Ala Ser Asn Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln

35 40 45

Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val Leu His His

50 55 60

Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe

65 70 75 80

Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe

85 90 95

Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly

100 105 110

Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp

115 120 125

Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu

130 135 140

Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly

145 150 155 160

Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala

165 170 175

His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln

180 185 190

Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu

195 200 205

Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala

210 215 220

Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser

225 230 235 240

Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro

245 250 255

Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly

260 265 270

Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly

275 280 285

Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys

290 295 300

Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr

305 310 315 320

Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser

325 330 335

Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro

340 345 350

Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr

355 360 365

Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val

370 375 380

Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp

385 390 395 400

Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro

405 410 415

Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu

420 425 430

Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser

435 440 445

Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile

450 455 460

Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu

465 470 475 480

Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg

485 490 495

Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu

500 505 510

Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp

515 520 525

Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala

530 535 540

Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His

545 550 555 560

Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn

565 570 575

Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val

580 585 590

Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn

595 600 605

Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile

610 615 620

Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala

625 630 635 640

Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser

645 650 655

Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu

660 665 670

Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe

675 680 685

Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr

690 695 700

Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His

705 710 715 720

Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val

725 730 735

Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser

740 745 750

Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp

755 760 765

Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr

770 775 780

Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser

785 790 795 800

Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys

805 810 815

Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys

820 825 830

Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala

835 840 845

Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly

850 855 860

Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu

865 870 875 880

His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile

885 890 895

Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr

900 905 910

Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu

915 920 925

Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp

930 935 940

Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn

945 950 955 960

Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser

965 970 975

His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met

980 985 990

Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser

995 1000 1005

Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys

1010 1015 1020

Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly

1025 1030 1035 1040

Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln

1045 1050 1055

Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu

1060 1065 1070

Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr

1075 1080 1085

Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser

1090 1095 1100

His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala

1105 1110 1115 1120

Gln Asp Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile

1125 1130 1135

Asp Tyr Lys Asp Asp Asp Asp Lys

1140

<210> 7

<211> 2295

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-3FT

<400> 7

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly

1140 1145 1150

Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser

1155 1160 1165

Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe

1170 1175 1180

Ser Thr Thr Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln Val

1185 1190 1195 1200

Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val Leu His His Val

1205 1210 1215

Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe Leu

1220 1225 1230

Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe Arg

1235 1240 1245

Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly Ile

1250 1255 1260

Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp Ser

1265 1270 1275 1280

Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu Asn

1285 1290 1295

Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly Gln

1300 1305 1310

Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala His

1315 1320 1325

Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln Leu

1330 1335 1340

Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu Val

1345 1350 1355 1360

Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala Gln

1365 1370 1375

Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser Ile

1380 1385 1390

Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro Trp

1395 1400 1405

Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly Ser

1410 1415 1420

Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly Phe

1425 1430 1435 1440

Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys Gly

1445 1450 1455

Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr Gln

1460 1465 1470

Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser Cys

1475 1480 1485

Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro Glu

1490 1495 1500

Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr Glu

1505 1510 1515 1520

Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val Lys

1525 1530 1535

Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp Val

1540 1545 1550

Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro Val

1555 1560 1565

Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu Gly

1570 1575 1580

Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser Lys

1585 1590 1595 1600

Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile Lys

1605 1610 1615

Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu Asn

1620 1625 1630

Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg Ser

1635 1640 1645

Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu Thr

1650 1655 1660

Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp Ala

1665 1670 1675 1680

Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala Ser

1685 1690 1695

Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His Leu

1700 1705 1710

Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn Phe

1715 1720 1725

Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val Gly

1730 1735 1740

Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn Leu

1745 1750 1755 1760

Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile Pro

1765 1770 1775

Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala Tyr

1780 1785 1790

Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser Ala

1795 1800 1805

Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu Arg

1810 1815 1820

Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe Cys

1825 1830 1835 1840

Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr Asp

1845 1850 1855

Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His Trp

1860 1865 1870

Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val Val

1875 1880 1885

Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser Gln

1890 1895 1900

Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp Asp

1905 1910 1915 1920

Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr Asp

1925 1930 1935

Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser Ser

1940 1945 1950

Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys Phe

1955 1960 1965

Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys Met

1970 1975 1980

Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala Gly

1985 1990 1995 2000

Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly Tyr

2005 2010 2015

Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu His

2020 2025 2030

Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile Glu

2035 2040 2045

Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr Ala

2050 2055 2060

Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu Ser

2065 2070 2075 2080

Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp Glu

2085 2090 2095

Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn Val

2100 2105 2110

Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser His

2115 2120 2125

Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met Val

2130 2135 2140

Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser Lys

2145 2150 2155 2160

Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys Asp

2165 2170 2175

Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly Asn

2180 2185 2190

Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln Phe

2195 2200 2205

Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu Ser

2210 2215 2220

His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr Thr

2225 2230 2235 2240

Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser His

2245 2250 2255

Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala Gln

2260 2265 2270

Asp Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp

2275 2280 2285

Tyr Lys Asp Asp Asp Asp Lys

2290 2295

<210> 8

<211> 1273

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-NpuN-3FT

<400> 8

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Leu Ser

1140 1145 1150

Tyr Glu Thr Glu Ile Leu Thr Val Glu Tyr Gly Leu Leu Pro Ile Gly

1155 1160 1165

Lys Ile Val Glu Lys Arg Ile Glu Cys Thr Val Tyr Ser Val Asp Asn

1170 1175 1180

Asn Gly Asn Ile Tyr Thr Gln Pro Val Ala Gln Trp His Asp Arg Gly

1185 1190 1195 1200

Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser Leu Ile Arg

1205 1210 1215

Ala Thr Lys Asp His Lys Phe Met Thr Val Asp Gly Gln Met Leu Pro

1220 1225 1230

Ile Asp Glu Ile Phe Glu Arg Glu Leu Asp Leu Met Arg Val Asp Asn

1235 1240 1245

Leu Pro Asn Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp

1250 1255 1260

Ile Asp Tyr Lys Asp Asp Asp Asp Lys

1265 1270

<210> 9

<211> 1182

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-NpuC-3FT

<400> 9

Met Ile Lys Ile Ala Thr Arg Lys Tyr Leu Gly Lys Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Glu Arg Asp His Asn Phe Ala Leu Lys Asn Gly Phe

20 25 30

Ile Ala Ser Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg

35 40 45

Lys Met Lys Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys

50 55 60

Ser Cys Ser Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp

65 70 75 80

Asp Leu Thr Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met

85 90 95

Asp Val Val Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile

100 105 110

Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg

115 120 125

Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu

130 135 140

Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe

145 150 155 160

Leu Lys Val Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly

165 170 175

Ala Gln Gln Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly

180 185 190

Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser

195 200 205

Pro Gly Ala Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro

210 215 220

Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln

225 230 235 240

His Val Gln Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser

245 250 255

His Lys Asp Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe

260 265 270

Leu Ala Leu Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro

275 280 285

Ala Leu Thr Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe

290 295 300

Ser Met Asp Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val

305 310 315 320

Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp

325 330 335

Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser

340 345 350

Val Ser Pro Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln

355 360 365

Val Asn Pro Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr

370 375 380

Met Leu Pro Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln

385 390 395 400

Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn

405 410 415

Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser

420 425 430

Leu Lys Ser Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser

435 440 445

Ile Gly Gly Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val

450 455 460

Gly Phe Leu Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro

465 470 475 480

Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu

485 490 495

Glu Thr Glu Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His

500 505 510

Ala Leu Val Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala

515 520 525

Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile

530 535 540

Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val

545 550 555 560

Leu Thr Thr Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser

565 570 575

Met Ser Phe Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg

580 585 590

Val Asn Lys Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr

595 600 605

Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val

610 615 620

Ser Ala Gly Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala

625 630 635 640

Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu

645 650 655

Tyr Gly Trp Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe

660 665 670

Asp Val Pro Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe

675 680 685

Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu

690 695 700

Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu

705 710 715 720

Ile Val Phe Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala

725 730 735

Leu Ser Gln Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His

740 745 750

Ser Ala Asn Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala

755 760 765

Met Val Val Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln

770 775 780

Arg His Phe Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro

785 790 795 800

Ile Val Asp Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile

805 810 815

Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys

820 825 830

Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly

835 840 845

Val Arg Pro Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly

850 855 860

Lys Thr Thr Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser

865 870 875 880

Gly Asp Ala Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu

885 890 895

Val His Gln Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu

900 905 910

Leu Leu Thr Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly

915 920 925

Val Pro Ala Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser

930 935 940

Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly

945 950 955 960

Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro

965 970 975

Pro Leu Val Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala

980 985 990

Arg Arg Met Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg

995 1000 1005

Ala Val Val Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys

1010 1015 1020

Thr Arg Leu Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr

1025 1030 1035 1040

Ile Gln His Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met

1045 1050 1055

Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val

1060 1065 1070

Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu Arg

1075 1080 1085

His Tyr Asn Met Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala Arg

1090 1095 1100

Ile Phe Gln Leu Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu Glu

1105 1110 1115 1120

Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe Ala

1125 1130 1135

Lys Gln Gln Thr Glu Ser His Asp Leu Pro Leu His Pro Arg Ala Ala

1140 1145 1150

Gly Ala Ser Arg Gln Ala Gln Asp Asp Tyr Lys Asp His Asp Gly Asp

1155 1160 1165

Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys

1170 1175 1180

<210> 10

<211> 1263

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-NpuN-3FT

<400> 10

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Leu Ser Tyr Glu Thr Glu Ile Leu Thr Val Glu Tyr

1140 1145 1150

Gly Leu Leu Pro Ile Gly Lys Ile Val Glu Lys Arg Ile Glu Cys Thr

1155 1160 1165

Val Tyr Ser Val Asp Asn Asn Gly Asn Ile Tyr Thr Gln Pro Val Ala

1170 1175 1180

Gln Trp His Asp Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1185 1190 1195 1200

Asp Gly Ser Leu Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Val

1205 1210 1215

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Glu Leu Asp

1220 1225 1230

Leu Met Arg Val Asp Asn Leu Pro Asn Asp Tyr Lys Asp His Asp Gly

1235 1240 1245

Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys

1250 1255 1260

<210> 11

<211> 1192

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-NpuC-3FT

<400> 11

Met Ile Lys Ile Ala Thr Arg Lys Tyr Leu Gly Lys Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Glu Arg Asp His Asn Phe Ala Leu Lys Asn Gly Phe

20 25 30

Ile Ala Ser Asn Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe

35 40 45

Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln

50 55 60

Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly

65 70 75 80

Phe Ser Thr Thr Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln

85 90 95

Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val Leu His His

100 105 110

Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe

115 120 125

Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe

130 135 140

Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly

145 150 155 160

Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp

165 170 175

Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu

180 185 190

Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly

195 200 205

Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala

210 215 220

His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln

225 230 235 240

Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu

245 250 255

Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala

260 265 270

Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser

275 280 285

Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro

290 295 300

Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly

305 310 315 320

Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly

325 330 335

Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys

340 345 350

Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr

355 360 365

Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser

370 375 380

Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro

385 390 395 400

Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr

405 410 415

Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val

420 425 430

Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp

435 440 445

Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro

450 455 460

Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu

465 470 475 480

Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser

485 490 495

Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile

500 505 510

Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu

515 520 525

Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg

530 535 540

Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu

545 550 555 560

Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp

565 570 575

Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala

580 585 590

Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His

595 600 605

Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn

610 615 620

Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val

625 630 635 640

Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn

645 650 655

Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile

660 665 670

Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala

675 680 685

Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser

690 695 700

Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu

705 710 715 720

Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe

725 730 735

Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr

740 745 750

Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His

755 760 765

Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val

770 775 780

Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser

785 790 795 800

Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp

805 810 815

Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr

820 825 830

Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser

835 840 845

Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys

850 855 860

Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys

865 870 875 880

Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala

885 890 895

Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly

900 905 910

Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu

915 920 925

His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile

930 935 940

Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr

945 950 955 960

Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu

965 970 975

Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp

980 985 990

Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn

995 1000 1005

Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser

1010 1015 1020

His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met

1025 1030 1035 1040

Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser

1045 1050 1055

Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys

1060 1065 1070

Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly

1075 1080 1085

Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln

1090 1095 1100

Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu

1105 1110 1115 1120

Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr

1125 1130 1135

Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser

1140 1145 1150

His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala

1155 1160 1165

Gln Asp Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile

1170 1175 1180

Asp Tyr Lys Asp Asp Asp Asp Lys

1185 1190

<210> 12

<211> 1311

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1188-NpuN-3FT

<400> 12

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly

1140 1145 1150

Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser

1155 1160 1165

Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe

1170 1175 1180

Ser Thr Thr Cys Leu Ser Tyr Glu Thr Glu Ile Leu Thr Val Glu Tyr

1185 1190 1195 1200

Gly Leu Leu Pro Ile Gly Lys Ile Val Glu Lys Arg Ile Glu Cys Thr

1205 1210 1215

Val Tyr Ser Val Asp Asn Asn Gly Asn Ile Tyr Thr Gln Pro Val Ala

1220 1225 1230

Gln Trp His Asp Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1235 1240 1245

Asp Gly Ser Leu Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Val

1250 1255 1260

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Glu Leu Asp

1265 1270 1275 1280

Leu Met Arg Val Asp Asn Leu Pro Asn Asp Tyr Lys Asp His Asp Gly

1285 1290 1295

Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys

1300 1305 1310

<210> 13

<211> 1144

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1188-NpuC-3FT

<400> 13

Met Ile Lys Ile Ala Thr Arg Lys Tyr Leu Gly Lys Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Glu Arg Asp His Asn Phe Ala Leu Lys Asn Gly Phe

20 25 30

Ile Ala Ser Asn Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln

35 40 45

Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val Leu His His

50 55 60

Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe

65 70 75 80

Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe

85 90 95

Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly

100 105 110

Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp

115 120 125

Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu

130 135 140

Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly

145 150 155 160

Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala

165 170 175

His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln

180 185 190

Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu

195 200 205

Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala

210 215 220

Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser

225 230 235 240

Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro

245 250 255

Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly

260 265 270

Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly

275 280 285

Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys

290 295 300

Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr

305 310 315 320

Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser

325 330 335

Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro

340 345 350

Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr

355 360 365

Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val

370 375 380

Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp

385 390 395 400

Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro

405 410 415

Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu

420 425 430

Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser

435 440 445

Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile

450 455 460

Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu

465 470 475 480

Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg

485 490 495

Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu

500 505 510

Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp

515 520 525

Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala

530 535 540

Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His

545 550 555 560

Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn

565 570 575

Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val

580 585 590

Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn

595 600 605

Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile

610 615 620

Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala

625 630 635 640

Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser

645 650 655

Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu

660 665 670

Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe

675 680 685

Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr

690 695 700

Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His

705 710 715 720

Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val

725 730 735

Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser

740 745 750

Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp

755 760 765

Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr

770 775 780

Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser

785 790 795 800

Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys

805 810 815

Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys

820 825 830

Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala

835 840 845

Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly

850 855 860

Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu

865 870 875 880

His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile

885 890 895

Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr

900 905 910

Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu

915 920 925

Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp

930 935 940

Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn

945 950 955 960

Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser

965 970 975

His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met

980 985 990

Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser

995 1000 1005

Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys

1010 1015 1020

Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly

1025 1030 1035 1040

Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln

1045 1050 1055

Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu

1060 1065 1070

Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr

1075 1080 1085

Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser

1090 1095 1100

His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala

1105 1110 1115 1120

Gln Asp Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile

1125 1130 1135

Asp Tyr Lys Asp Asp Asp Asp Lys

1140

<210> 14

<211> 179

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP-71- CfaN

<400> 14

Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu

1 5 10 15

Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly

20 25 30

Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile

35 40 45

Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr

50 55 60

Leu Thr Tyr Gly Val Gln Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr

65 70 75 80

Val Glu Tyr Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile

85 90 95

Glu Cys Thr Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln

100 105 110

Pro Ile Ala Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr

115 120 125

Cys Leu Glu Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe

130 135 140

Met Thr Thr Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg

145 150 155 160

Gly Leu Asp Leu Lys Gln Val Asp Gly Leu Pro Gly His His His His

165 170 175

His His Gly

<210> 15

<211> 213

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP-71- CfaC

<400> 15

Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu Lys Asn Gly Leu

20 25 30

Val Ala Ser Asn Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His

35 40 45

Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr

50 55 60

Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys

65 70 75 80

Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp

85 90 95

Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr

100 105 110

Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile

115 120 125

Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln

130 135 140

Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val

145 150 155 160

Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys

165 170 175

Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr

180 185 190

Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys Gly His His

195 200 205

His His His His Gly

210

<210> 16

<211> 247

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP

<400> 16

Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu

1 5 10 15

Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly

20 25 30

Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile

35 40 45

Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr

50 55 60

Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys

65 70 75 80

Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu

85 90 95

Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu

100 105 110

Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly

115 120 125

Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr

130 135 140

Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn

145 150 155 160

Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser

165 170 175

Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly

180 185 190

Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu

195 200 205

Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe

210 215 220

Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys Gly

225 230 235 240

His His His His His His Gly

245

<210> 17

<211> 180

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP-71- NpuN

<400> 17

Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu

1 5 10 15

Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly

20 25 30

Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile

35 40 45

Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr

50 55 60

Leu Thr Tyr Gly Val Gln Cys Leu Ser Tyr Glu Thr Glu Ile Leu Thr

65 70 75 80

Val Glu Tyr Gly Leu Leu Pro Ile Gly Lys Ile Val Glu Lys Arg Ile

85 90 95

Glu Cys Thr Val Tyr Ser Val Asp Asn Asn Gly Asn Ile Tyr Thr Gln

100 105 110

Pro Val Ala Gln Trp His Asp Arg Gly Glu Gln Glu Val Phe Glu Tyr

115 120 125

Cys Leu Glu Asp Gly Ser Leu Ile Arg Ala Thr Lys Asp His Lys Phe

130 135 140

Met Thr Val Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg

145 150 155 160

Glu Leu Asp Leu Met Arg Val Asp Asn Leu Pro Asn Gly His His His

165 170 175

His His His Gly

180

<210> 18

<211> 213

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP-71- NpuC

<400> 18

Met Ile Lys Ile Ala Thr Arg Lys Tyr Leu Gly Lys Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Glu Arg Asp His Asn Phe Ala Leu Lys Asn Gly Phe

20 25 30

Ile Ala Ser Asn Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His

35 40 45

Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr

50 55 60

Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys

65 70 75 80

Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp

85 90 95

Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr

100 105 110

Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile

115 120 125

Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln

130 135 140

Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val

145 150 155 160

Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys

165 170 175

Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr

180 185 190

Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys Gly His His

195 200 205

His His His His Gly

210

<210> 19

<211> 1371

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-CfaN-3FT-SopE

<400> 19

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Leu Ser

1140 1145 1150

Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Phe Leu Pro Ile Gly

1155 1160 1165

Lys Ile Val Glu Glu Arg Ile Glu Cys Thr Val Tyr Thr Val Asp Lys

1170 1175 1180

Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala Gln Trp His Asn Arg Gly

1185 1190 1195 1200

Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser Ile Ile Arg

1205 1210 1215

Ala Thr Lys Asp His Lys Phe Met Thr Thr Asp Gly Gln Met Leu Pro

1220 1225 1230

Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp Leu Lys Gln Val Asp Gly

1235 1240 1245

Leu Pro Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile

1250 1255 1260

Asp Tyr Lys Asp Asp Asp Asp Lys Thr Lys Ile Thr Leu Ser Pro Gln

1265 1270 1275 1280

Asn Phe Arg Ile Gln Lys Gln Glu Thr Thr Leu Leu Lys Glu Lys Ser

1285 1290 1295

Thr Glu Lys Asn Ser Leu Ala Lys Ser Ile Leu Ala Val Lys Asn His

1300 1305 1310

Phe Ile Glu Leu Arg Ser Lys Leu Ser Glu Arg Phe Ile Ser His Lys

1315 1320 1325

Asn Thr Glu Ser Ser Ala Thr His Phe His Arg Gly Ser Ala Ser Glu

1330 1335 1340

Gly Arg Ala Val Leu Thr Asn Lys Val Val Lys Asp Phe Met Leu Gln

1345 1350 1355 1360

Thr Leu Asn Asp Ile Asp Ile Arg Gly Ser Ala

1365 1370

<210> 20

<211> 1281

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-CfaC-3FT-SopE

<400> 20

Met Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys Gln

1 5 10 15

Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu Ala

20 25 30

Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser Lys

35 40 45

Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala Thr

50 55 60

His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu Thr Asn

65 70 75 80

Lys Val Val Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile Asp Ile

85 90 95

Arg Gly Ser Ala Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln

100 105 110

Asn Val Tyr Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu Lys

115 120 125

Asn Gly Leu Val Ala Ser Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr

130 135 140

Leu Val Arg Lys Met Lys Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu

145 150 155 160

Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala

165 170 175

His Val Asp Asp Leu Thr Pro Glu Gln Val Leu Asp Gly Asp Val Asn

180 185 190

Glu Leu Met Asp Val Val Leu His His Val Pro Glu Ala Lys Leu Val

195 200 205

Glu Cys Ile Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe

210 215 220

Lys His Arg Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu

225 230 235 240

Ala Asp Leu Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu

245 250 255

Glu Ile Phe Leu Lys Val Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe

260 265 270

Ala Gly Gly Ala Gln Gln Lys Arg Glu Asn Val Asn Pro Arg His Pro

275 280 285

Cys Leu Gly Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn

290 295 300

Val Cys Ser Pro Gly Ala Pro Ala Ala His Pro Glu Gly Gln Pro Pro

305 310 315 320

Pro Glu Pro Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu

325 330 335

Val Leu Gln His Val Gln Ala Leu Leu Val Lys Arg Phe Gln His Thr

340 345 350

Ile Arg Ser His Lys Asp Phe Leu Ala Gln Ile Val Leu Pro Ala Thr

355 360 365

Phe Val Phe Leu Ala Leu Met Leu Ser Ile Val Ile Pro Pro Phe Gly

370 375 380

Glu Tyr Pro Ala Leu Thr Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr

385 390 395 400

Thr Phe Phe Ser Met Asp Glu Pro Gly Ser Glu Gln Phe Thr Val Leu

405 410 415

Ala Asp Val Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys

420 425 430

Glu Gly Trp Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys

435 440 445

Thr Pro Ser Val Ser Pro Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys

450 455 460

Trp Thr Gln Val Asn Pro Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu

465 470 475 480

Lys Leu Thr Met Leu Pro Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro

485 490 495

Pro Pro Gln Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr

500 505 510

Asp Arg Asn Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile

515 520 525

Arg Ser Ser Leu Lys Ser Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly

530 535 540

Gly Ile Ser Ile Gly Gly Lys Leu Pro Val Val Pro Ile Thr Gly Glu

545 550 555 560

Ala Leu Val Gly Phe Leu Ser Asp Leu Gly Arg Ile Met Asn Val Ser

565 570 575

Gly Gly Pro Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu

580 585 590

Lys His Leu Glu Thr Glu Asp Asn Ile Lys Val Trp Phe Asn Asn Lys

595 600 605

Gly Trp His Ala Leu Val Ser Phe Leu Asn Val Ala His Asn Ala Ile

610 615 620

Leu Arg Ala Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile

625 630 635 640

Thr Val Ile Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu

645 650 655

Ile Thr Val Leu Thr Thr Ser Val Asp Ala Val Val Ala Ile Cys Val

660 665 670

Ile Phe Ser Met Ser Phe Val Pro Ala Ser Phe Val Leu Tyr Leu Ile

675 680 685

Gln Glu Arg Val Asn Lys Ser Lys His Leu Gln Phe Ile Ser Gly Val

690 695 700

Ser Pro Thr Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp Ile Met Asn

705 710 715 720

Tyr Ser Val Ser Ala Gly Leu Val Val Gly Ile Phe Ile Gly Phe Gln

725 730 735

Lys Lys Ala Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu Val Ala Leu

740 745 750

Leu Leu Leu Tyr Gly Trp Ala Val Ile Pro Met Met Tyr Pro Ala Ser

755 760 765

Phe Leu Phe Asp Val Pro Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala

770 775 780

Asn Leu Phe Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu

785 790 795 800

Leu Phe Glu Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala Val Leu Arg

805 810 815

Lys Leu Leu Ile Val Phe Pro His Phe Cys Leu Gly Arg Gly Leu Ile

820 825 830

Asp Leu Ala Leu Ser Gln Ala Val Thr Asp Val Tyr Ala Arg Phe Gly

835 840 845

Glu Glu His Ser Ala Asn Pro Phe His Trp Asp Leu Ile Gly Lys Asn

850 855 860

Leu Phe Ala Met Val Val Glu Gly Val Val Tyr Phe Leu Leu Thr Leu

865 870 875 880

Leu Val Gln Arg His Phe Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr

885 890 895

Lys Glu Pro Ile Val Asp Glu Asp Asp Asp Val Ala Glu Glu Arg Gln

900 905 910

Arg Ile Ile Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg Leu His Glu

915 920 925

Leu Thr Lys Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val Asp Arg Leu

930 935 940

Cys Val Gly Val Arg Pro Gly Glu Cys Phe Gly Leu Leu Gly Val Asn

945 950 955 960

Gly Ala Gly Lys Thr Thr Thr Phe Lys Met Leu Thr Gly Asp Thr Thr

965 970 975

Val Thr Ser Gly Asp Ala Thr Val Ala Gly Lys Ser Ile Leu Thr Asn

980 985 990

Ile Ser Glu Val His Gln Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala

995 1000 1005

Ile Asp Glu Leu Leu Thr Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg

1010 1015 1020

Leu Arg Gly Val Pro Ala Glu Glu Ile Glu Lys Val Ala Asn Trp Ser

1025 1030 1035 1040

Ile Lys Ser Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr

1045 1050 1055

Tyr Ser Gly Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile

1060 1065 1070

Gly Cys Pro Pro Leu Val Leu Leu Asp Glu Pro Thr Thr Gly Met Asp

1075 1080 1085

Pro Gln Ala Arg Arg Met Leu Trp Asn Val Ile Val Ser Ile Ile Arg

1090 1095 1100

Glu Gly Arg Ala Val Val Leu Thr Ser His Ser Met Glu Glu Cys Glu

1105 1110 1115 1120

Ala Leu Cys Thr Arg Leu Ala Ile Met Val Lys Gly Ala Phe Arg Cys

1125 1130 1135

Met Gly Thr Ile Gln His Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile

1140 1145 1150

Val Thr Met Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu

1155 1160 1165

Asn Pro Val Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly Ser Val Gln

1170 1175 1180

Arg Glu Arg His Tyr Asn Met Leu Gln Phe Gln Val Ser Ser Ser Ser

1185 1190 1195 1200

Leu Ala Arg Ile Phe Gln Leu Leu Leu Ser His Lys Asp Ser Leu Leu

1205 1210 1215

Ile Glu Glu Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln Val Phe Val

1220 1225 1230

Asn Phe Ala Lys Gln Gln Thr Glu Ser His Asp Leu Pro Leu His Pro

1235 1240 1245

Arg Ala Ala Gly Ala Ser Arg Gln Ala Gln Asp Asp Tyr Lys Asp His

1250 1255 1260

Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp Asp Asp Asp

1265 1270 1275 1280

Lys

<210> 21

<211> 1361

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaN-3FT-SopE

<400> 21

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr

1140 1145 1150

Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr

1155 1160 1165

Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala

1170 1175 1180

Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1185 1190 1195 1200

Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr

1205 1210 1215

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp

1220 1225 1230

Leu Lys Gln Val Asp Gly Leu Pro Asp Tyr Lys Asp His Asp Gly Asp

1235 1240 1245

Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys Thr Lys

1250 1255 1260

Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys Gln Glu Thr Thr

1265 1270 1275 1280

Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu Ala Lys Ser Ile

1285 1290 1295

Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser Lys Leu Ser Glu

1300 1305 1310

Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala Thr His Phe His

1315 1320 1325

Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu Thr Asn Lys Val Val

1330 1335 1340

Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile Asp Ile Arg Gly Ser

1345 1350 1355 1360

Ala

<210> 22

<211> 1291

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaC-3FT-SopE

<400> 22

Met Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys Gln

1 5 10 15

Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu Ala

20 25 30

Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser Lys

35 40 45

Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala Thr

50 55 60

His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu Thr Asn

65 70 75 80

Lys Val Val Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile Asp Ile

85 90 95

Arg Gly Ser Ala Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln

100 105 110

Asn Val Tyr Asp Ile Gly Val Gly Glu Pro His Asn Phe Leu Leu Lys

115 120 125

Asn Gly Leu Val Ala Ser Asn Cys Ser Gly Thr Pro Leu Phe Leu Lys

130 135 140

Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys

145 150 155 160

Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser

165 170 175

Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp Asp Leu Thr

180 185 190

Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val

195 200 205

Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu

210 215 220

Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala

225 230 235 240

Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser

245 250 255

Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val

260 265 270

Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln

275 280 285

Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu

290 295 300

Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala

305 310 315 320

Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro

325 330 335

Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln

340 345 350

Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp

355 360 365

Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu

370 375 380

Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr

385 390 395 400

Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp

405 410 415

Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn

420 425 430

Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu

435 440 445

Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro

450 455 460

Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro

465 470 475 480

Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro

485 490 495

Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln

500 505 510

Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp

515 520 525

Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser

530 535 540

Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly

545 550 555 560

Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu

565 570 575

Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg

580 585 590

Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu

595 600 605

Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val

610 615 620

Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro

625 630 635 640

Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro

645 650 655

Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr

660 665 670

Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe

675 680 685

Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys

690 695 700

Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp

705 710 715 720

Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly

725 730 735

Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser

740 745 750

Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp

755 760 765

Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro

770 775 780

Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile

785 790 795 800

Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg

805 810 815

Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe

820 825 830

Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln

835 840 845

Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn

850 855 860

Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val

865 870 875 880

Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe

885 890 895

Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp

900 905 910

Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly

915 920 925

Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro

930 935 940

Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro

945 950 955 960

Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr

965 970 975

Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala

980 985 990

Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln

995 1000 1005

Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr

1010 1015 1020

Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala

1025 1030 1035 1040

Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu

1045 1050 1055

Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys

1060 1065 1070

Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val

1075 1080 1085

Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met

1090 1095 1100

Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val

1105 1110 1115 1120

Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu

1125 1130 1135

Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His

1140 1145 1150

Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys

1155 1160 1165

Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe

1170 1175 1180

Phe Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn

1185 1190 1195 1200

Met Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln

1205 1210 1215

Leu Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val

1220 1225 1230

Thr Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln

1235 1240 1245

Thr Glu Ser His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser

1250 1255 1260

Arg Gln Ala Gln Asp Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp

1265 1270 1275 1280

His Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys

1285 1290

<210> 23

<211> 278

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP-71-CfaN-SopE

<400> 23

Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu

1 5 10 15

Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly

20 25 30

Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile

35 40 45

Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr

50 55 60

Leu Thr Tyr Gly Val Gln Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr

65 70 75 80

Val Glu Tyr Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile

85 90 95

Glu Cys Thr Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln

100 105 110

Pro Ile Ala Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr

115 120 125

Cys Leu Glu Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe

130 135 140

Met Thr Thr Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg

145 150 155 160

Gly Leu Asp Leu Lys Gln Val Asp Gly Leu Pro Gly His His His His

165 170 175

His His Gly Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln

180 185 190

Lys Gln Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser

195 200 205

Leu Ala Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg

210 215 220

Ser Lys Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser

225 230 235 240

Ala Thr His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu

245 250 255

Thr Asn Lys Val Val Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile

260 265 270

Asp Ile Arg Gly Ser Ala

275

<210> 24

<211> 312

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP-71-CfaC-SopE

<400> 24

Met Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys Gln

1 5 10 15

Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu Ala

20 25 30

Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser Lys

35 40 45

Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala Thr

50 55 60

His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu Thr Asn

65 70 75 80

Lys Val Val Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile Asp Ile

85 90 95

Arg Gly Ser Ala Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln

100 105 110

Asn Val Tyr Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu Lys

115 120 125

Asn Gly Leu Val Ala Ser Asn Cys Phe Ser Arg Tyr Pro Asp His Met

130 135 140

Lys Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln

145 150 155 160

Glu Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala

165 170 175

Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys

180 185 190

Gly Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu

195 200 205

Tyr Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys

210 215 220

Asn Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly

225 230 235 240

Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp

245 250 255

Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala

260 265 270

Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu

275 280 285

Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys

290 295 300

Gly His His His His His His Gly

305 310

<210> 25

<211> 192

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP-71-CfaN-DD1

<400> 25

Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu

1 5 10 15

Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly

20 25 30

Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile

35 40 45

Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr

50 55 60

Leu Thr Tyr Gly Val Gln Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr

65 70 75 80

Val Glu Tyr Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile

85 90 95

Glu Cys Thr Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln

100 105 110

Pro Ile Ala Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr

115 120 125

Cys Leu Glu Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe

130 135 140

Met Thr Thr Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg

145 150 155 160

Gly Leu Asp Leu Lys Gln Val Asp Gly Leu Pro Gly His His His His

165 170 175

His His Gly Arg Ile Ser Phe Gly Ser Pro Pro Pro Met Ala Gly Gly

180 185 190

<210> 26

<211> 226

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> EGFP-71-CfaC-DD1

<400> 26

Met Arg Ile Ser Phe Gly Ser Pro Pro Pro Met Ala Gly Gly Val Lys

1 5 10 15

Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr Asp Ile Gly

20 25 30

Val Glu Lys Asp His Asn Phe Leu Leu Lys Asn Gly Leu Val Ala Ser

35 40 45

Asn Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe

50 55 60

Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe

65 70 75 80

Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly

85 90 95

Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu

100 105 110

Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His

115 120 125

Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile Lys Val Asn

130 135 140

Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp

145 150 155 160

His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro

165 170 175

Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn

180 185 190

Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly

195 200 205

Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys Gly His His His His His

210 215 220

His Gly

225

<210> 27

<211> 101

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> CfaN

<400> 27

Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Phe Leu

1 5 10 15

Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr Val Tyr Thr

20 25 30

Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala Gln Trp His

35 40 45

Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser

50 55 60

Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr Asp Gly Gln

65 70 75 80

Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp Leu Lys Gln

85 90 95

Val Asp Gly Leu Pro

100

<210> 28

<211> 36

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> CfaC

<400> 28

Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu Lys Asn Gly Leu

20 25 30

Val Ala Ser Asn

35

<210> 29

<211> 36

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> CfaCmut

<400> 29

Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Gly Glu Pro His Asn Phe Leu Leu Lys Asn Gly Leu

20 25 30

Val Ala Ser Asn

35

<210> 30

<211> 30

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> CatN

<400> 30

Cys Leu Ser Gly Asp Thr Met Ile Glu Ile Leu Asp Asp Asp Gly Ile

1 5 10 15

Ile Gln Lys Ile Ser Met Glu Asp Leu Tyr Gln Arg Leu Ala

20 25 30

<210> 31

<211> 104

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> CatC

<400> 31

Met Phe Lys Leu Asn Thr Lys Asn Ile Lys Val Leu Thr Pro Ser Gly

1 5 10 15

Phe Lys Ser Phe Ser Gly Ile Gln Lys Val Tyr Lys Pro Phe Tyr His

20 25 30

His Ile Ile Phe Asp Asp Gly Ser Glu Ile Lys Cys Ser Asp Asn His

35 40 45

Ser Phe Gly Lys Asp Lys Ile Lys Ala Ser Thr Ile Lys Val Gly Asp

50 55 60

Tyr Leu Gln Gly Lys Lys Val Leu Tyr Asn Glu Ile Val Glu Glu Gly

65 70 75 80

Ile Tyr Leu Tyr Asp Leu Leu Asn Val Gly Glu Asp Asn Leu Tyr Tyr

85 90 95

Thr Asn Gly Ile Val Ser His Asn

100

<210> 32

<211> 102

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> NpuN

<400> 32

Cys Leu Ser Tyr Glu Thr Glu Ile Leu Thr Val Glu Tyr Gly Leu Leu

1 5 10 15

Pro Ile Gly Lys Ile Val Glu Lys Arg Ile Glu Cys Thr Val Tyr Ser

20 25 30

Val Asp Asn Asn Gly Asn Ile Tyr Thr Gln Pro Val Ala Gln Trp His

35 40 45

Asp Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser

50 55 60

Leu Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Val Asp Gly Gln

65 70 75 80

Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Glu Leu Asp Leu Met Arg

85 90 95

Val Asp Asn Leu Pro Asn

100

<210> 33

<211> 36

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> NpuC

<400> 33

Met Ile Lys Ile Ala Thr Arg Lys Tyr Leu Gly Lys Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Glu Arg Asp His Asn Phe Ala Leu Lys Asn Gly Phe

20 25 30

Ile Ala Ser Asn

35

<210> 34

<211> 28

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> M3

<400> 34

Lys Ser Val Thr Leu Glu Ser Arg Ser Pro Lys Phe Leu Asn Trp Phe

1 5 10 15

Ser Val Phe Ser Leu Phe Lys Val Ile Thr Thr Gly

20 25

<210> 35

<211> 21

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> M4

<400> 35

Tyr Met Ser Ile Leu Arg Cys Ala Ser Gly Lys Ile Ser Ile Ala Ala

1 5 10 15

Pro Pro Tyr Ile Phe

20

<210> 36

<211> 36

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> NpuCmut

<400> 36

Met Ile Lys Ile Ala Thr Arg Lys Tyr Leu Gly Lys Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Gly Glu Pro His Asn Phe Ala Leu Lys Asn Gly Phe

20 25 30

Ile Ala Ser Asn

35

<210> 37

<211> 34

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> M5

<400> 37

Ala Gly Glu Ser Phe Asn Phe Met Val Lys Leu Leu Tyr Lys His Pro

1 5 10 15

Ile Leu Pro Cys Leu Lys Thr Leu Leu Ser Ile Arg Ser Ser Cys Ser

20 25 30

Pro Arg

<210> 38

<211> 88

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> GP41N

<400> 38

Cys Leu Asp Leu Lys Thr Gln Val Gln Thr Pro Gln Gly Met Lys Glu

1 5 10 15

Ile Ser Asn Ile Gln Val Gly Asp Leu Val Leu Ser Asn Thr Gly Tyr

20 25 30

Asn Glu Val Leu Asn Val Phe Pro Lys Ser Lys Lys Lys Ser Tyr Lys

35 40 45

Ile Thr Leu Glu Asp Gly Lys Glu Ile Ile Cys Ser Glu Glu His Leu

50 55 60

Phe Pro Thr Gln Thr Gly Glu Met Asn Ile Ser Gly Gly Leu Lys Glu

65 70 75 80

Gly Met Cys Leu Tyr Val Lys Glu

85

<210> 39

<211> 102

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ConN

<400> 39

Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Ala Val

1 5 10 15

Pro Ile Gly Lys Ile Val Glu Glu Asn Ile Glu Cys Thr Val Tyr Ser

20 25 30

Val Asp Glu Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala Gln Trp His

35 40 45

Asp Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser

50 55 60

Thr Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Glu Asp Gly Glu

65 70 75 80

Met Leu Pro Ile Asp Glu Ile Phe Glu Gln Gly Leu Asp Leu Lys Gln

85 90 95

Val Lys Gly Leu Pro Asp

100

<210> 40

<211> 16

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> CL1

<400> 40

Ala Cys Lys Asn Trp Phe Ser Ser Leu Ser His Phe Val Ile His Leu

1 5 10 15

<210> 41

<211> 62

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> Deg1

<400> 41

Met Asn Lys Ile Pro Ile Lys Asp Leu Leu Asn Pro Gln Ile Thr Asp

1 5 10 15

Glu Phe Lys Ser Ser Ile Leu Asp Ile Asn Lys Lys Leu Phe Ser Ile

20 25 30

Cys Cys Asn Leu Pro Lys Leu Pro Glu Ser Val Thr Thr Glu Glu Glu

35 40 45

Val Glu Leu Arg Asp Ile Leu Gly Phe Leu Ser Arg Ala Asn

50 55 60

<210> 42

<211> 30

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> PEST

<400> 42

Arg Ser Ser Ser Pro Ser Asp Ser Asp Thr Ser Gly Phe Ser Ser Gly

1 5 10 15

Ser Asp His Leu Ser Asp Leu Ile Ser Ser Leu Arg Ile Ser

20 25 30

<210> 43

<211> 13

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> DD1

<220>

<221> 变体

<222> 12

<223> X= 任意氨基酸，优选Gly

<400> 43

Arg Ile Ser Phe Gly Ser Pro Pro Pro Met Ala Xaa Gly

1 5 10

<210> 44

<211> 9

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> DD2

<220>

<221> 变体

<222> 8

<223> X= 任意氨基酸，优选Gly

<400> 44

Gly Ser Pro Pro Pro Met Ala Xaa Gly

1 5

<210> 45

<211> 11

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> DD3

<400> 45

Thr Asn Gly Ile Leu Lys Leu Gly Cys Gln Gly

1 5 10

<210> 46

<211> 17

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> M1

<400> 46

Cys Ser Glu Ile Ile Pro Met Ser Arg Ser Thr Pro Ile Ser Thr Met

1 5 10 15

Gly

<210> 47

<211> 19

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> M2

<400> 47

Val Ser Phe Ala Phe Asn Leu Asn Ser Leu Ile Val Gly Ile Leu Arg

1 5 10 15

Phe His Trp

<210> 48

<211> 100

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> SopE

<400> 48

Met Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys Gln

1 5 10 15

Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu Ala

20 25 30

Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser Lys

35 40 45

Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala Thr

50 55 60

His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu Thr Asn

65 70 75 80

Lys Val Val Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile Asp Ile

85 90 95

Arg Gly Ser Ala

100

<210> 49

<211> 78

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> SopE-1-78

<400> 49

Met Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys Gln

1 5 10 15

Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu Ala

20 25 30

Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser Lys

35 40 45

Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala Thr

50 55 60

His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu

65 70 75

<210> 50

<211> 63

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> SopE-15-78

<400> 50

Gln Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu

1 5 10 15

Ala Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser

20 25 30

Lys Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala

35 40 45

Thr His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu

50 55 60

<210> 51

<211> 35

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> SopE-15-50

<400> 51

Gln Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu

1 5 10 15

Ala Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser

20 25 30

Lys Leu Ser

35

<210> 52

<211> 35

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> L2

<400> 52

Ser Leu Ile Ser Leu Pro Leu Pro Thr Arg Val Lys Phe Ser Ser Leu

1 5 10 15

Leu Leu Ile Arg Ile Met Lys Ile Ile Thr Met Thr Phe Pro Lys Lys

20 25 30

Leu Arg Ser

35

<210> 53

<211> 16

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> L6

<400> 53

Phe Tyr Tyr Pro Ile Trp Phe Ala Arg Val Leu Leu Val His Tyr Gln

1 5 10 15

<210> 54

<211> 46

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> L9

<400> 54

Ser Asn Pro Phe Ser Ser Leu Phe Gly Ala Ser Leu Leu Ile Asp Ser

1 5 10 15

Val Ser Leu Lys Ser Asn Trp Asp Thr Ser Ser Ser Ser Cys Leu Ile

20 25 30

Ser Phe Phe Ser Ser Val Met Phe Ser Ser Thr Thr Arg Ser

35 40 45

<210> 55

<211> 39

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> L10

<400> 55

Cys Arg Gln Arg Phe Ser Cys His Leu Thr Ala Ser Tyr Pro Gln Ser

1 5 10 15

Thr Val Thr Pro Phe Leu Ala Phe Leu Arg Arg Asp Phe Phe Phe Leu

20 25 30

Arg His Asn Ser Ser Ala Asp

35

<210> 56

<211> 46

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> L11

<400> 56

Gly Ala Pro His Val Val Leu Phe Asp Phe Glu Leu Arg Ile Thr Asn

1 5 10 15

Pro Leu Ser His Ile Gln Ser Val Ser Leu Gln Ile Thr Leu Ile Phe

20 25 30

Cys Ser Leu Pro Ser Leu Ile Leu Ser Lys Phe Leu Gln Val

35 40 45

<210> 57

<211> 39

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> L12

<400> 57

Asn Thr Pro Leu Phe Ser Lys Ser Phe Ser Thr Thr Cys Gly Val Ala

1 5 10 15

Lys Lys Thr Leu Leu Leu Ala Gln Ile Ser Ser Leu Phe Phe Leu Leu

20 25 30

Leu Ser Ser Asn Ile Ala Val

35

<210> 58

<211> 45

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> L15

<400> 58

Pro Thr Val Lys Asn Ser Pro Lys Ile Phe Cys Leu Ser Ser Ser Pro

1 5 10 15

Tyr Leu Ala Phe Asn Leu Glu Tyr Leu Ser Leu Arg Ile Phe Ser Thr

20 25 30

Leu Ser Lys Cys Ser Asn Thr Leu Leu Thr Ser Leu Ser

35 40 45

<210> 59

<211> 30

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> L16

<400> 59

Ser Asn Gln Leu Lys Arg Leu Trp Leu Trp Leu Leu Glu Val Arg Ser

1 5 10 15

Phe Asp Arg Thr Leu Arg Arg Pro Trp Ile His Leu Pro Ser

20 25 30

<210> 60

<211> 1149

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4N-1150

<400> 60

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn

1140 1145

<210> 61

<211> 1124

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4C-1150

<400> 61

Cys Phe Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn

1 5 10 15

Ile Gln Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser

20 25 30

Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp Asp Leu Thr Pro

35 40 45

Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val Leu

50 55 60

His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu

65 70 75 80

Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser

85 90 95

Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser

100 105 110

Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr

115 120 125

Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys

130 135 140

Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys

145 150 155 160

Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro

165 170 175

Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly

180 185 190

Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln Ala

195 200 205

Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp Phe

210 215 220

Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met

225 230 235 240

Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu

245 250 255

His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu

260 265 270

Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys

275 280 285

Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr

290 295 300

Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn

305 310 315 320

Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser

325 330 335

Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu

340 345 350

Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg

355 360 365

Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe

370 375 380

Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys

385 390 395 400

Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys

405 410 415

Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser

420 425 430

Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu

435 440 445

Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp

450 455 460

Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser

465 470 475 480

Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys

485 490 495

Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu

500 505 510

Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser

515 520 525

Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val

530 535 540

Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser

545 550 555 560

Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val

565 570 575

Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu

580 585 590

Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro

595 600 605

Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala

610 615 620

Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser

625 630 635 640

Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn

645 650 655

Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr

660 665 670

Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro

675 680 685

His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala

690 695 700

Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro

705 710 715 720

Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu

725 730 735

Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe

740 745 750

Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu

755 760 765

Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn

770 775 780

Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly

785 790 795 800

Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly

805 810 815

Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr

820 825 830

Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr

835 840 845

Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn

850 855 860

Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly

865 870 875 880

Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu

885 890 895

Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr

900 905 910

Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg

915 920 925

Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu

930 935 940

Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu

945 950 955 960

Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu

965 970 975

Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala

980 985 990

Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu

995 1000 1005

Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser

1010 1015 1020

Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe

1025 1030 1035 1040

Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met

1045 1050 1055

Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu

1060 1065 1070

Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr

1075 1080 1085

Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr

1090 1095 1100

Glu Ser His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg

1105 1110 1115 1120

Gln Ala Gln Asp

<210> 62

<211> 1139

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4N-1140

<400> 62

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr

<210> 63

<211> 1134

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4C-1140

<400> 63

Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly Thr Gly Leu

1 5 10 15

Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser Gln Arg Lys

20 25 30

Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe Ser Thr Thr

35 40 45

Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln Val Leu Asp Gly

50 55 60

Asp Val Asn Glu Leu Met Asp Val Val Leu His His Val Pro Glu Ala

65 70 75 80

Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn

85 90 95

Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu

100 105 110

Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr

115 120 125

Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp Ser Asp Ser Gly

130 135 140

Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu Asn Val Asn Pro

145 150 155 160

Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln

165 170 175

Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala His Pro Glu Gly

180 185 190

Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly

195 200 205

Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu Val Lys Arg Phe

210 215 220

Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala Gln Ile Val Leu

225 230 235 240

Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser Ile Val Ile Pro

245 250 255

Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro Trp Ile Tyr Gly

260 265 270

Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly Ser Glu Gln Phe

275 280 285

Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg

290 295 300

Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr

305 310 315 320

Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr Gln Leu Phe Gln

325 330 335

Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser Cys Arg Cys Ser

340 345 350

Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro Glu Gly Ala Gly

355 360 365

Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln

370 375 380

Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro

385 390 395 400

Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp Val Asn Glu Gln

405 410 415

Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro Val Val Pro Ile

420 425 430

Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu Gly Arg Ile Met

435 440 445

Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro

450 455 460

Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile Lys Val Trp Phe

465 470 475 480

Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu Asn Val Ala His

485 490 495

Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu

500 505 510

Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln

515 520 525

Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp Ala Val Val Ala

530 535 540

Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala Ser Phe Val Leu

545 550 555 560

Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His Leu Gln Phe Ile

565 570 575

Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp

580 585 590

Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val Gly Ile Phe Ile

595 600 605

Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu

610 615 620

Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile Pro Met Met Tyr

625 630 635 640

Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala Tyr Val Ala Leu

645 650 655

Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe

660 665 670

Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala

675 680 685

Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe Cys Leu Gly Arg

690 695 700

Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr Asp Val Tyr Ala

705 710 715 720

Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His Trp Asp Leu Ile

725 730 735

Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val Val Tyr Phe Leu

740 745 750

Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser Gln Trp Ile Ala

755 760 765

Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp Asp Val Ala Glu

770 775 780

Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg

785 790 795 800

Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val

805 810 815

Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys Phe Gly Leu Leu

820 825 830

Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys Met Leu Thr Gly

835 840 845

Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala Gly Lys Ser Ile

850 855 860

Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly Tyr Cys Pro Gln

865 870 875 880

Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu His Leu Tyr Leu

885 890 895

Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile Glu Lys Val Ala

900 905 910

Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu

915 920 925

Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile

930 935 940

Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp Glu Pro Thr Thr

945 950 955 960

Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn Val Ile Val Ser

965 970 975

Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser His Ser Met Glu

980 985 990

Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met Val Lys Gly Ala

995 1000 1005

Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser Lys Phe Gly Asp

1010 1015 1020

Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu

1025 1030 1035 1040

Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly

1045 1050 1055

Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln Phe Gln Val Ser

1060 1065 1070

Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu Ser His Lys Asp

1075 1080 1085

Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln

1090 1095 1100

Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser His Asp Leu Pro

1105 1110 1115 1120

Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala Gln Asp

1125 1130

<210> 64

<211> 1187

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4N-1188

<400> 64

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly

1140 1145 1150

Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser

1155 1160 1165

Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe

1170 1175 1180

Ser Thr Thr

1185

<210> 65

<211> 1086

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4C-1188

<400> 65

Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln Val Leu Asp Gly

1 5 10 15

Asp Val Asn Glu Leu Met Asp Val Val Leu His His Val Pro Glu Ala

20 25 30

Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn

35 40 45

Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu

50 55 60

Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr

65 70 75 80

Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp Ser Asp Ser Gly

85 90 95

Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu Asn Val Asn Pro

100 105 110

Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln

115 120 125

Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala His Pro Glu Gly

130 135 140

Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly

145 150 155 160

Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu Val Lys Arg Phe

165 170 175

Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala Gln Ile Val Leu

180 185 190

Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser Ile Val Ile Pro

195 200 205

Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro Trp Ile Tyr Gly

210 215 220

Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly Ser Glu Gln Phe

225 230 235 240

Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg

245 250 255

Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr

260 265 270

Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr Gln Leu Phe Gln

275 280 285

Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser Cys Arg Cys Ser

290 295 300

Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro Glu Gly Ala Gly

305 310 315 320

Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln

325 330 335

Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro

340 345 350

Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp Val Asn Glu Gln

355 360 365

Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro Val Val Pro Ile

370 375 380

Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu Gly Arg Ile Met

385 390 395 400

Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro

405 410 415

Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile Lys Val Trp Phe

420 425 430

Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu Asn Val Ala His

435 440 445

Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu

450 455 460

Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln

465 470 475 480

Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp Ala Val Val Ala

485 490 495

Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala Ser Phe Val Leu

500 505 510

Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His Leu Gln Phe Ile

515 520 525

Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp

530 535 540

Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val Gly Ile Phe Ile

545 550 555 560

Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu

565 570 575

Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile Pro Met Met Tyr

580 585 590

Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala Tyr Val Ala Leu

595 600 605

Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe

610 615 620

Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala

625 630 635 640

Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe Cys Leu Gly Arg

645 650 655

Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr Asp Val Tyr Ala

660 665 670

Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His Trp Asp Leu Ile

675 680 685

Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val Val Tyr Phe Leu

690 695 700

Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser Gln Trp Ile Ala

705 710 715 720

Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp Asp Val Ala Glu

725 730 735

Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg

740 745 750

Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val

755 760 765

Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys Phe Gly Leu Leu

770 775 780

Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys Met Leu Thr Gly

785 790 795 800

Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala Gly Lys Ser Ile

805 810 815

Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly Tyr Cys Pro Gln

820 825 830

Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu His Leu Tyr Leu

835 840 845

Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile Glu Lys Val Ala

850 855 860

Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu

865 870 875 880

Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile

885 890 895

Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp Glu Pro Thr Thr

900 905 910

Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn Val Ile Val Ser

915 920 925

Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser His Ser Met Glu

930 935 940

Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met Val Lys Gly Ala

945 950 955 960

Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser Lys Phe Gly Asp

965 970 975

Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu

980 985 990

Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly

995 1000 1005

Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln Phe Gln Val Ser

1010 1015 1020

Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu Ser His Lys Asp

1025 1030 1035 1040

Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln

1045 1050 1055

Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser His Asp Leu Pro

1060 1065 1070

Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala Gln Asp

1075 1080 1085

<210> 66

<211> 1250

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-CfaN

<400> 66

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Leu Ser

1140 1145 1150

Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Phe Leu Pro Ile Gly

1155 1160 1165

Lys Ile Val Glu Glu Arg Ile Glu Cys Thr Val Tyr Thr Val Asp Lys

1170 1175 1180

Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala Gln Trp His Asn Arg Gly

1185 1190 1195 1200

Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser Ile Ile Arg

1205 1210 1215

Ala Thr Lys Asp His Lys Phe Met Thr Thr Asp Gly Gln Met Leu Pro

1220 1225 1230

Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp Leu Lys Gln Val Asp Gly

1235 1240 1245

Leu Pro

1250

<210> 67

<211> 1160

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-CfaC

<400> 67

Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu Lys Asn Gly Leu

20 25 30

Val Ala Ser Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg

35 40 45

Lys Met Lys Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys

50 55 60

Ser Cys Ser Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp

65 70 75 80

Asp Leu Thr Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met

85 90 95

Asp Val Val Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile

100 105 110

Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg

115 120 125

Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu

130 135 140

Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe

145 150 155 160

Leu Lys Val Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly

165 170 175

Ala Gln Gln Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly

180 185 190

Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser

195 200 205

Pro Gly Ala Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro

210 215 220

Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln

225 230 235 240

His Val Gln Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser

245 250 255

His Lys Asp Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe

260 265 270

Leu Ala Leu Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro

275 280 285

Ala Leu Thr Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe

290 295 300

Ser Met Asp Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val

305 310 315 320

Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp

325 330 335

Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser

340 345 350

Val Ser Pro Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln

355 360 365

Val Asn Pro Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr

370 375 380

Met Leu Pro Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln

385 390 395 400

Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn

405 410 415

Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser

420 425 430

Leu Lys Ser Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser

435 440 445

Ile Gly Gly Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val

450 455 460

Gly Phe Leu Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro

465 470 475 480

Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu

485 490 495

Glu Thr Glu Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His

500 505 510

Ala Leu Val Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala

515 520 525

Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile

530 535 540

Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val

545 550 555 560

Leu Thr Thr Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser

565 570 575

Met Ser Phe Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg

580 585 590

Val Asn Lys Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr

595 600 605

Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val

610 615 620

Ser Ala Gly Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala

625 630 635 640

Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu

645 650 655

Tyr Gly Trp Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe

660 665 670

Asp Val Pro Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe

675 680 685

Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu

690 695 700

Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu

705 710 715 720

Ile Val Phe Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala

725 730 735

Leu Ser Gln Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His

740 745 750

Ser Ala Asn Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala

755 760 765

Met Val Val Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln

770 775 780

Arg His Phe Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro

785 790 795 800

Ile Val Asp Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile

805 810 815

Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys

820 825 830

Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly

835 840 845

Val Arg Pro Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly

850 855 860

Lys Thr Thr Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser

865 870 875 880

Gly Asp Ala Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu

885 890 895

Val His Gln Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu

900 905 910

Leu Leu Thr Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly

915 920 925

Val Pro Ala Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser

930 935 940

Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly

945 950 955 960

Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro

965 970 975

Pro Leu Val Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala

980 985 990

Arg Arg Met Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg

995 1000 1005

Ala Val Val Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys

1010 1015 1020

Thr Arg Leu Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr

1025 1030 1035 1040

Ile Gln His Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met

1045 1050 1055

Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val

1060 1065 1070

Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu Arg

1075 1080 1085

His Tyr Asn Met Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala Arg

1090 1095 1100

Ile Phe Gln Leu Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu Glu

1105 1110 1115 1120

Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe Ala

1125 1130 1135

Lys Gln Gln Thr Glu Ser His Asp Leu Pro Leu His Pro Arg Ala Ala

1140 1145 1150

Gly Ala Ser Arg Gln Ala Gln Asp

1155 1160

<210> 68

<211> 1240

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaN

<400> 68

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr

1140 1145 1150

Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr

1155 1160 1165

Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala

1170 1175 1180

Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1185 1190 1195 1200

Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr

1205 1210 1215

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp

1220 1225 1230

Leu Lys Gln Val Asp Gly Leu Pro

1235 1240

<210> 69

<211> 1170

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaCmut

<400> 69

Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Gly Glu Pro His Asn Phe Leu Leu Lys Asn Gly Leu

20 25 30

Val Ala Ser Asn Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe

35 40 45

Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln

50 55 60

Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly

65 70 75 80

Phe Ser Thr Thr Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln

85 90 95

Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val Leu His His

100 105 110

Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe

115 120 125

Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe

130 135 140

Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly

145 150 155 160

Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp

165 170 175

Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu

180 185 190

Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly

195 200 205

Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala

210 215 220

His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln

225 230 235 240

Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu

245 250 255

Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala

260 265 270

Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser

275 280 285

Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro

290 295 300

Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly

305 310 315 320

Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly

325 330 335

Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys

340 345 350

Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr

355 360 365

Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser

370 375 380

Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro

385 390 395 400

Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr

405 410 415

Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val

420 425 430

Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp

435 440 445

Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro

450 455 460

Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu

465 470 475 480

Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser

485 490 495

Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile

500 505 510

Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu

515 520 525

Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg

530 535 540

Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu

545 550 555 560

Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp

565 570 575

Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala

580 585 590

Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His

595 600 605

Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn

610 615 620

Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val

625 630 635 640

Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn

645 650 655

Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile

660 665 670

Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala

675 680 685

Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser

690 695 700

Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu

705 710 715 720

Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe

725 730 735

Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr

740 745 750

Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His

755 760 765

Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val

770 775 780

Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser

785 790 795 800

Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp

805 810 815

Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr

820 825 830

Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser

835 840 845

Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys

850 855 860

Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys

865 870 875 880

Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala

885 890 895

Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly

900 905 910

Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu

915 920 925

His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile

930 935 940

Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr

945 950 955 960

Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu

965 970 975

Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp

980 985 990

Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn

995 1000 1005

Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser

1010 1015 1020

His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met

1025 1030 1035 1040

Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser

1045 1050 1055

Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys

1060 1065 1070

Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly

1075 1080 1085

Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln

1090 1095 1100

Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu

1105 1110 1115 1120

Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr

1125 1130 1135

Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser

1140 1145 1150

His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala

1155 1160 1165

Gln Asp

1170

<210> 70

<211> 1288

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1188-CfaN

<400> 70

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly

1140 1145 1150

Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser

1155 1160 1165

Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe

1170 1175 1180

Ser Thr Thr Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr

1185 1190 1195 1200

Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr

1205 1210 1215

Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala

1220 1225 1230

Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1235 1240 1245

Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr

1250 1255 1260

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp

1265 1270 1275 1280

Leu Lys Gln Val Asp Gly Leu Pro

1285

<210> 71

<211> 1122

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1188-Cfacmut

<400> 71

Met Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr

1 5 10 15

Asp Ile Gly Val Gly Glu Pro His Asn Phe Leu Leu Lys Asn Gly Leu

20 25 30

Val Ala Ser Asn Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln

35 40 45

Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val Leu His His

50 55 60

Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe

65 70 75 80

Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe

85 90 95

Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly

100 105 110

Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp

115 120 125

Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu

130 135 140

Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly

145 150 155 160

Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala

165 170 175

His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln

180 185 190

Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu

195 200 205

Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala

210 215 220

Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser

225 230 235 240

Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro

245 250 255

Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly

260 265 270

Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly

275 280 285

Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys

290 295 300

Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr

305 310 315 320

Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser

325 330 335

Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro

340 345 350

Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr

355 360 365

Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val

370 375 380

Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp

385 390 395 400

Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro

405 410 415

Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu

420 425 430

Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser

435 440 445

Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile

450 455 460

Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu

465 470 475 480

Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg

485 490 495

Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu

500 505 510

Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp

515 520 525

Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala

530 535 540

Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His

545 550 555 560

Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn

565 570 575

Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val

580 585 590

Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn

595 600 605

Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile

610 615 620

Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala

625 630 635 640

Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser

645 650 655

Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu

660 665 670

Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe

675 680 685

Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr

690 695 700

Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His

705 710 715 720

Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val

725 730 735

Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser

740 745 750

Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp

755 760 765

Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr

770 775 780

Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser

785 790 795 800

Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys

805 810 815

Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys

820 825 830

Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala

835 840 845

Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly

850 855 860

Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu

865 870 875 880

His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile

885 890 895

Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr

900 905 910

Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu

915 920 925

Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp

930 935 940

Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn

945 950 955 960

Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser

965 970 975

His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met

980 985 990

Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser

995 1000 1005

Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys

1010 1015 1020

Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly

1025 1030 1035 1040

Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln

1045 1050 1055

Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu

1060 1065 1070

Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr

1075 1080 1085

Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser

1090 1095 1100

His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala

1105 1110 1115 1120

Gln Asp

<210> 72

<211> 1349

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-CfaN-SopE

<400> 72

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Leu Ser

1140 1145 1150

Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Phe Leu Pro Ile Gly

1155 1160 1165

Lys Ile Val Glu Glu Arg Ile Glu Cys Thr Val Tyr Thr Val Asp Lys

1170 1175 1180

Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala Gln Trp His Asn Arg Gly

1185 1190 1195 1200

Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser Ile Ile Arg

1205 1210 1215

Ala Thr Lys Asp His Lys Phe Met Thr Thr Asp Gly Gln Met Leu Pro

1220 1225 1230

Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp Leu Lys Gln Val Asp Gly

1235 1240 1245

Leu Pro Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys

1250 1255 1260

Gln Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu

1265 1270 1275 1280

Ala Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser

1285 1290 1295

Lys Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala

1300 1305 1310

Thr His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu Thr

1315 1320 1325

Asn Lys Val Val Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile Asp

1330 1335 1340

Ile Arg Gly Ser Ala

1345

<210> 73

<211> 1259

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-CfaC-SopE

<400> 73

Met Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys Gln

1 5 10 15

Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu Ala

20 25 30

Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser Lys

35 40 45

Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala Thr

50 55 60

His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu Thr Asn

65 70 75 80

Lys Val Val Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile Asp Ile

85 90 95

Arg Gly Ser Ala Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln

100 105 110

Asn Val Tyr Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu Lys

115 120 125

Asn Gly Leu Val Ala Ser Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr

130 135 140

Leu Val Arg Lys Met Lys Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu

145 150 155 160

Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala

165 170 175

His Val Asp Asp Leu Thr Pro Glu Gln Val Leu Asp Gly Asp Val Asn

180 185 190

Glu Leu Met Asp Val Val Leu His His Val Pro Glu Ala Lys Leu Val

195 200 205

Glu Cys Ile Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe

210 215 220

Lys His Arg Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu

225 230 235 240

Ala Asp Leu Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu

245 250 255

Glu Ile Phe Leu Lys Val Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe

260 265 270

Ala Gly Gly Ala Gln Gln Lys Arg Glu Asn Val Asn Pro Arg His Pro

275 280 285

Cys Leu Gly Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn

290 295 300

Val Cys Ser Pro Gly Ala Pro Ala Ala His Pro Glu Gly Gln Pro Pro

305 310 315 320

Pro Glu Pro Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu

325 330 335

Val Leu Gln His Val Gln Ala Leu Leu Val Lys Arg Phe Gln His Thr

340 345 350

Ile Arg Ser His Lys Asp Phe Leu Ala Gln Ile Val Leu Pro Ala Thr

355 360 365

Phe Val Phe Leu Ala Leu Met Leu Ser Ile Val Ile Pro Pro Phe Gly

370 375 380

Glu Tyr Pro Ala Leu Thr Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr

385 390 395 400

Thr Phe Phe Ser Met Asp Glu Pro Gly Ser Glu Gln Phe Thr Val Leu

405 410 415

Ala Asp Val Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys

420 425 430

Glu Gly Trp Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys

435 440 445

Thr Pro Ser Val Ser Pro Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys

450 455 460

Trp Thr Gln Val Asn Pro Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu

465 470 475 480

Lys Leu Thr Met Leu Pro Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro

485 490 495

Pro Pro Gln Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr

500 505 510

Asp Arg Asn Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile

515 520 525

Arg Ser Ser Leu Lys Ser Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly

530 535 540

Gly Ile Ser Ile Gly Gly Lys Leu Pro Val Val Pro Ile Thr Gly Glu

545 550 555 560

Ala Leu Val Gly Phe Leu Ser Asp Leu Gly Arg Ile Met Asn Val Ser

565 570 575

Gly Gly Pro Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu

580 585 590

Lys His Leu Glu Thr Glu Asp Asn Ile Lys Val Trp Phe Asn Asn Lys

595 600 605

Gly Trp His Ala Leu Val Ser Phe Leu Asn Val Ala His Asn Ala Ile

610 615 620

Leu Arg Ala Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile

625 630 635 640

Thr Val Ile Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu

645 650 655

Ile Thr Val Leu Thr Thr Ser Val Asp Ala Val Val Ala Ile Cys Val

660 665 670

Ile Phe Ser Met Ser Phe Val Pro Ala Ser Phe Val Leu Tyr Leu Ile

675 680 685

Gln Glu Arg Val Asn Lys Ser Lys His Leu Gln Phe Ile Ser Gly Val

690 695 700

Ser Pro Thr Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp Ile Met Asn

705 710 715 720

Tyr Ser Val Ser Ala Gly Leu Val Val Gly Ile Phe Ile Gly Phe Gln

725 730 735

Lys Lys Ala Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu Val Ala Leu

740 745 750

Leu Leu Leu Tyr Gly Trp Ala Val Ile Pro Met Met Tyr Pro Ala Ser

755 760 765

Phe Leu Phe Asp Val Pro Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala

770 775 780

Asn Leu Phe Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu

785 790 795 800

Leu Phe Glu Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala Val Leu Arg

805 810 815

Lys Leu Leu Ile Val Phe Pro His Phe Cys Leu Gly Arg Gly Leu Ile

820 825 830

Asp Leu Ala Leu Ser Gln Ala Val Thr Asp Val Tyr Ala Arg Phe Gly

835 840 845

Glu Glu His Ser Ala Asn Pro Phe His Trp Asp Leu Ile Gly Lys Asn

850 855 860

Leu Phe Ala Met Val Val Glu Gly Val Val Tyr Phe Leu Leu Thr Leu

865 870 875 880

Leu Val Gln Arg His Phe Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr

885 890 895

Lys Glu Pro Ile Val Asp Glu Asp Asp Asp Val Ala Glu Glu Arg Gln

900 905 910

Arg Ile Ile Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg Leu His Glu

915 920 925

Leu Thr Lys Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val Asp Arg Leu

930 935 940

Cys Val Gly Val Arg Pro Gly Glu Cys Phe Gly Leu Leu Gly Val Asn

945 950 955 960

Gly Ala Gly Lys Thr Thr Thr Phe Lys Met Leu Thr Gly Asp Thr Thr

965 970 975

Val Thr Ser Gly Asp Ala Thr Val Ala Gly Lys Ser Ile Leu Thr Asn

980 985 990

Ile Ser Glu Val His Gln Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala

995 1000 1005

Ile Asp Glu Leu Leu Thr Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg

1010 1015 1020

Leu Arg Gly Val Pro Ala Glu Glu Ile Glu Lys Val Ala Asn Trp Ser

1025 1030 1035 1040

Ile Lys Ser Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr

1045 1050 1055

Tyr Ser Gly Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile

1060 1065 1070

Gly Cys Pro Pro Leu Val Leu Leu Asp Glu Pro Thr Thr Gly Met Asp

1075 1080 1085

Pro Gln Ala Arg Arg Met Leu Trp Asn Val Ile Val Ser Ile Ile Arg

1090 1095 1100

Glu Gly Arg Ala Val Val Leu Thr Ser His Ser Met Glu Glu Cys Glu

1105 1110 1115 1120

Ala Leu Cys Thr Arg Leu Ala Ile Met Val Lys Gly Ala Phe Arg Cys

1125 1130 1135

Met Gly Thr Ile Gln His Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile

1140 1145 1150

Val Thr Met Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu

1155 1160 1165

Asn Pro Val Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly Ser Val Gln

1170 1175 1180

Arg Glu Arg His Tyr Asn Met Leu Gln Phe Gln Val Ser Ser Ser Ser

1185 1190 1195 1200

Leu Ala Arg Ile Phe Gln Leu Leu Leu Ser His Lys Asp Ser Leu Leu

1205 1210 1215

Ile Glu Glu Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln Val Phe Val

1220 1225 1230

Asn Phe Ala Lys Gln Gln Thr Glu Ser His Asp Leu Pro Leu His Pro

1235 1240 1245

Arg Ala Ala Gly Ala Ser Arg Gln Ala Gln Asp

1250 1255

<210> 74

<211> 1339

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaN-SopE

<400> 74

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr

1140 1145 1150

Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr

1155 1160 1165

Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala

1170 1175 1180

Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1185 1190 1195 1200

Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr

1205 1210 1215

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp

1220 1225 1230

Leu Lys Gln Val Asp Gly Leu Pro Thr Lys Ile Thr Leu Ser Pro Gln

1235 1240 1245

Asn Phe Arg Ile Gln Lys Gln Glu Thr Thr Leu Leu Lys Glu Lys Ser

1250 1255 1260

Thr Glu Lys Asn Ser Leu Ala Lys Ser Ile Leu Ala Val Lys Asn His

1265 1270 1275 1280

Phe Ile Glu Leu Arg Ser Lys Leu Ser Glu Arg Phe Ile Ser His Lys

1285 1290 1295

Asn Thr Glu Ser Ser Ala Thr His Phe His Arg Gly Ser Ala Ser Glu

1300 1305 1310

Gly Arg Ala Val Leu Thr Asn Lys Val Val Lys Asp Phe Met Leu Gln

1315 1320 1325

Thr Leu Asn Asp Ile Asp Ile Arg Gly Ser Ala

1330 1335

<210> 75

<211> 1269

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaCmut-SopE

<400> 75

Met Thr Lys Ile Thr Leu Ser Pro Gln Asn Phe Arg Ile Gln Lys Gln

1 5 10 15

Glu Thr Thr Leu Leu Lys Glu Lys Ser Thr Glu Lys Asn Ser Leu Ala

20 25 30

Lys Ser Ile Leu Ala Val Lys Asn His Phe Ile Glu Leu Arg Ser Lys

35 40 45

Leu Ser Glu Arg Phe Ile Ser His Lys Asn Thr Glu Ser Ser Ala Thr

50 55 60

His Phe His Arg Gly Ser Ala Ser Glu Gly Arg Ala Val Leu Thr Asn

65 70 75 80

Lys Val Val Lys Asp Phe Met Leu Gln Thr Leu Asn Asp Ile Asp Ile

85 90 95

Arg Gly Ser Ala Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln

100 105 110

Asn Val Tyr Asp Ile Gly Val Gly Glu Pro His Asn Phe Leu Leu Lys

115 120 125

Asn Gly Leu Val Ala Ser Asn Cys Ser Gly Thr Pro Leu Phe Leu Lys

130 135 140

Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys

145 150 155 160

Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser

165 170 175

Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp Asp Leu Thr

180 185 190

Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val

195 200 205

Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu

210 215 220

Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala

225 230 235 240

Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser

245 250 255

Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val

260 265 270

Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln

275 280 285

Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu

290 295 300

Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala

305 310 315 320

Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro

325 330 335

Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln

340 345 350

Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp

355 360 365

Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu

370 375 380

Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr

385 390 395 400

Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp

405 410 415

Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn

420 425 430

Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu

435 440 445

Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro

450 455 460

Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro

465 470 475 480

Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro

485 490 495

Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln

500 505 510

Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp

515 520 525

Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser

530 535 540

Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly

545 550 555 560

Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu

565 570 575

Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg

580 585 590

Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu

595 600 605

Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val

610 615 620

Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro

625 630 635 640

Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro

645 650 655

Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr

660 665 670

Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe

675 680 685

Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys

690 695 700

Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp

705 710 715 720

Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly

725 730 735

Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser

740 745 750

Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp

755 760 765

Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro

770 775 780

Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile

785 790 795 800

Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg

805 810 815

Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe

820 825 830

Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln

835 840 845

Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn

850 855 860

Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val

865 870 875 880

Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe

885 890 895

Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp

900 905 910

Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly

915 920 925

Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro

930 935 940

Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro

945 950 955 960

Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr

965 970 975

Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala

980 985 990

Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln

995 1000 1005

Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr

1010 1015 1020

Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala

1025 1030 1035 1040

Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu

1045 1050 1055

Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys

1060 1065 1070

Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val

1075 1080 1085

Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met

1090 1095 1100

Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val

1105 1110 1115 1120

Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu

1125 1130 1135

Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His

1140 1145 1150

Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys

1155 1160 1165

Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe

1170 1175 1180

Phe Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn

1185 1190 1195 1200

Met Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln

1205 1210 1215

Leu Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val

1220 1225 1230

Thr Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln

1235 1240 1245

Thr Glu Ser His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser

1250 1255 1260

Arg Gln Ala Gln Asp

1265

<210> 76

<211> 1263

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-CfaN-DD1

<400> 76

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Leu Ser

1140 1145 1150

Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Phe Leu Pro Ile Gly

1155 1160 1165

Lys Ile Val Glu Glu Arg Ile Glu Cys Thr Val Tyr Thr Val Asp Lys

1170 1175 1180

Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala Gln Trp His Asn Arg Gly

1185 1190 1195 1200

Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser Ile Ile Arg

1205 1210 1215

Ala Thr Lys Asp His Lys Phe Met Thr Thr Asp Gly Gln Met Leu Pro

1220 1225 1230

Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp Leu Lys Gln Val Asp Gly

1235 1240 1245

Leu Pro Arg Ile Ser Phe Gly Ser Pro Pro Pro Met Ala Gly Gly

1250 1255 1260

<210> 77

<211> 1173

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1150-CfaC-DD1

<400> 77

Met Arg Ile Ser Phe Gly Ser Pro Pro Pro Met Ala Gly Gly Val Lys

1 5 10 15

Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr Asp Ile Gly

20 25 30

Val Glu Lys Asp His Asn Phe Leu Leu Lys Asn Gly Leu Val Ala Ser

35 40 45

Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys

50 55 60

Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser

65 70 75 80

Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp Asp Leu Thr

85 90 95

Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp Val Val

100 105 110

Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly Gln Glu

115 120 125

Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala Tyr Ala

130 135 140

Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser

145 150 155 160

Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu Lys Val

165 170 175

Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln

180 185 190

Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro Arg Glu

195 200 205

Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro Gly Ala

210 215 220

Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro

225 230 235 240

Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His Val Gln

245 250 255

Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser His Lys Asp

260 265 270

Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu Ala Leu

275 280 285

Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr

290 295 300

Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp

305 310 315 320

Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu Leu Asn

325 330 335

Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu

340 345 350

Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val Ser Pro

355 360 365

Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val Asn Pro

370 375 380

Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met Leu Pro

385 390 395 400

Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln

405 410 415

Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp

420 425 430

Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser

435 440 445

Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly

450 455 460

Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly Phe Leu

465 470 475 480

Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile Thr Arg

485 490 495

Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu Thr Glu

500 505 510

Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala Leu Val

515 520 525

Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser Leu Pro

530 535 540

Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro

545 550 555 560

Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu Thr Thr

565 570 575

Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser Met Ser Phe

580 585 590

Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys

595 600 605

Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp

610 615 620

Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser Ala Gly

625 630 635 640

Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser

645 650 655

Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp

660 665 670

Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro

675 680 685

Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile

690 695 700

Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg

705 710 715 720

Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile Val Phe

725 730 735

Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln

740 745 750

Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser Ala Asn

755 760 765

Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met Val Val

770 775 780

Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg His Phe

785 790 795 800

Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile Val Asp

805 810 815

Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly

820 825 830

Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile Tyr Pro

835 840 845

Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val Arg Pro

850 855 860

Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys Thr Thr

865 870 875 880

Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly Asp Ala

885 890 895

Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val His Gln

900 905 910

Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr

915 920 925

Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala

930 935 940

Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu

945 950 955 960

Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys

965 970 975

Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro Leu Val

980 985 990

Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg Arg Met

995 1000 1005

Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala Val Val

1010 1015 1020

Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu

1025 1030 1035 1040

Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile Gln His

1045 1050 1055

Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys Ile Lys

1060 1065 1070

Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu Gln Phe

1075 1080 1085

Phe Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu Arg His Tyr Asn

1090 1095 1100

Met Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln

1105 1110 1115 1120

Leu Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val

1125 1130 1135

Thr Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe Ala Lys Gln Gln

1140 1145 1150

Thr Glu Ser His Asp Leu Pro Leu His Pro Arg Ala Ala Gly Ala Ser

1155 1160 1165

Arg Gln Ala Gln Asp

1170

<210> 78

<211> 1253

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaN-DD1

<400> 78

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr

1140 1145 1150

Gly Phe Leu Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr

1155 1160 1165

Val Tyr Thr Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala

1170 1175 1180

Gln Trp His Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu

1185 1190 1195 1200

Asp Gly Ser Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr

1205 1210 1215

Asp Gly Gln Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp

1220 1225 1230

Leu Lys Gln Val Asp Gly Leu Pro Arg Ile Ser Phe Gly Ser Pro Pro

1235 1240 1245

Pro Met Ala Gly Gly

1250

<210> 79

<211> 1183

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4-1140-CfaCmut-DD1

<400> 79

Met Arg Ile Ser Phe Gly Ser Pro Pro Pro Met Ala Gly Gly Val Lys

1 5 10 15

Ile Ile Ser Arg Lys Ser Leu Gly Thr Gln Asn Val Tyr Asp Ile Gly

20 25 30

Val Gly Glu Pro His Asn Phe Leu Leu Lys Asn Gly Leu Val Ala Ser

35 40 45

Asn Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly Thr Gly

50 55 60

Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser Gln Arg

65 70 75 80

Lys Gly Ser Glu Gly Thr Cys Ser Cys Ser Ser Lys Gly Phe Ser Thr

85 90 95

Thr Cys Pro Ala His Val Asp Asp Leu Thr Pro Glu Gln Val Leu Asp

100 105 110

Gly Asp Val Asn Glu Leu Met Asp Val Val Leu His His Val Pro Glu

115 120 125

Ala Lys Leu Val Glu Cys Ile Gly Gln Glu Leu Ile Phe Leu Leu Pro

130 135 140

Asn Lys Asn Phe Lys His Arg Ala Tyr Ala Ser Leu Phe Arg Glu Leu

145 150 155 160

Glu Glu Thr Leu Ala Asp Leu Gly Leu Ser Ser Phe Gly Ile Ser Asp

165 170 175

Thr Pro Leu Glu Glu Ile Phe Leu Lys Val Thr Glu Asp Ser Asp Ser

180 185 190

Gly Pro Leu Phe Ala Gly Gly Ala Gln Gln Lys Arg Glu Asn Val Asn

195 200 205

Pro Arg His Pro Cys Leu Gly Pro Arg Glu Lys Ala Gly Gln Thr Pro

210 215 220

Gln Asp Ser Asn Val Cys Ser Pro Gly Ala Pro Ala Ala His Pro Glu

225 230 235 240

Gly Gln Pro Pro Pro Glu Pro Glu Cys Pro Gly Pro Gln Leu Asn Thr

245 250 255

Gly Thr Gln Leu Val Leu Gln His Val Gln Ala Leu Leu Val Lys Arg

260 265 270

Phe Gln His Thr Ile Arg Ser His Lys Asp Phe Leu Ala Gln Ile Val

275 280 285

Leu Pro Ala Thr Phe Val Phe Leu Ala Leu Met Leu Ser Ile Val Ile

290 295 300

Pro Pro Phe Gly Glu Tyr Pro Ala Leu Thr Leu His Pro Trp Ile Tyr

305 310 315 320

Gly Gln Gln Tyr Thr Phe Phe Ser Met Asp Glu Pro Gly Ser Glu Gln

325 330 335

Phe Thr Val Leu Ala Asp Val Leu Leu Asn Lys Pro Gly Phe Gly Asn

340 345 350

Arg Cys Leu Lys Glu Gly Trp Leu Pro Glu Tyr Pro Cys Gly Asn Ser

355 360 365

Thr Pro Trp Lys Thr Pro Ser Val Ser Pro Asn Ile Thr Gln Leu Phe

370 375 380

Gln Lys Gln Lys Trp Thr Gln Val Asn Pro Ser Pro Ser Cys Arg Cys

385 390 395 400

Ser Thr Arg Glu Lys Leu Thr Met Leu Pro Glu Cys Pro Glu Gly Ala

405 410 415

Gly Gly Leu Pro Pro Pro Gln Arg Thr Gln Arg Ser Thr Glu Ile Leu

420 425 430

Gln Asp Leu Thr Asp Arg Asn Ile Ser Asp Phe Leu Val Lys Thr Tyr

435 440 445

Pro Ala Leu Ile Arg Ser Ser Leu Lys Ser Lys Phe Trp Val Asn Glu

450 455 460

Gln Arg Tyr Gly Gly Ile Ser Ile Gly Gly Lys Leu Pro Val Val Pro

465 470 475 480

Ile Thr Gly Glu Ala Leu Val Gly Phe Leu Ser Asp Leu Gly Arg Ile

485 490 495

Met Asn Val Ser Gly Gly Pro Ile Thr Arg Glu Ala Ser Lys Glu Ile

500 505 510

Pro Asp Phe Leu Lys His Leu Glu Thr Glu Asp Asn Ile Lys Val Trp

515 520 525

Phe Asn Asn Lys Gly Trp His Ala Leu Val Ser Phe Leu Asn Val Ala

530 535 540

His Asn Ala Ile Leu Arg Ala Ser Leu Pro Lys Asp Arg Ser Pro Glu

545 550 555 560

Glu Tyr Gly Ile Thr Val Ile Ser Gln Pro Leu Asn Leu Thr Lys Glu

565 570 575

Gln Leu Ser Glu Ile Thr Val Leu Thr Thr Ser Val Asp Ala Val Val

580 585 590

Ala Ile Cys Val Ile Phe Ser Met Ser Phe Val Pro Ala Ser Phe Val

595 600 605

Leu Tyr Leu Ile Gln Glu Arg Val Asn Lys Ser Lys His Leu Gln Phe

610 615 620

Ile Ser Gly Val Ser Pro Thr Thr Tyr Trp Val Thr Asn Phe Leu Trp

625 630 635 640

Asp Ile Met Asn Tyr Ser Val Ser Ala Gly Leu Val Val Gly Ile Phe

645 650 655

Ile Gly Phe Gln Lys Lys Ala Tyr Thr Ser Pro Glu Asn Leu Pro Ala

660 665 670

Leu Val Ala Leu Leu Leu Leu Tyr Gly Trp Ala Val Ile Pro Met Met

675 680 685

Tyr Pro Ala Ser Phe Leu Phe Asp Val Pro Ser Thr Ala Tyr Val Ala

690 695 700

Leu Ser Cys Ala Asn Leu Phe Ile Gly Ile Asn Ser Ser Ala Ile Thr

705 710 715 720

Phe Ile Leu Glu Leu Phe Glu Asn Asn Arg Thr Leu Leu Arg Phe Asn

725 730 735

Ala Val Leu Arg Lys Leu Leu Ile Val Phe Pro His Phe Cys Leu Gly

740 745 750

Arg Gly Leu Ile Asp Leu Ala Leu Ser Gln Ala Val Thr Asp Val Tyr

755 760 765

Ala Arg Phe Gly Glu Glu His Ser Ala Asn Pro Phe His Trp Asp Leu

770 775 780

Ile Gly Lys Asn Leu Phe Ala Met Val Val Glu Gly Val Val Tyr Phe

785 790 795 800

Leu Leu Thr Leu Leu Val Gln Arg His Phe Phe Leu Ser Gln Trp Ile

805 810 815

Ala Glu Pro Thr Lys Glu Pro Ile Val Asp Glu Asp Asp Asp Val Ala

820 825 830

Glu Glu Arg Gln Arg Ile Ile Thr Gly Gly Asn Lys Thr Asp Ile Leu

835 840 845

Arg Leu His Glu Leu Thr Lys Ile Tyr Pro Gly Thr Ser Ser Pro Ala

850 855 860

Val Asp Arg Leu Cys Val Gly Val Arg Pro Gly Glu Cys Phe Gly Leu

865 870 875 880

Leu Gly Val Asn Gly Ala Gly Lys Thr Thr Thr Phe Lys Met Leu Thr

885 890 895

Gly Asp Thr Thr Val Thr Ser Gly Asp Ala Thr Val Ala Gly Lys Ser

900 905 910

Ile Leu Thr Asn Ile Ser Glu Val His Gln Asn Met Gly Tyr Cys Pro

915 920 925

Gln Phe Asp Ala Ile Asp Glu Leu Leu Thr Gly Arg Glu His Leu Tyr

930 935 940

Leu Tyr Ala Arg Leu Arg Gly Val Pro Ala Glu Glu Ile Glu Lys Val

945 950 955 960

Ala Asn Trp Ser Ile Lys Ser Leu Gly Leu Thr Val Tyr Ala Asp Cys

965 970 975

Leu Ala Gly Thr Tyr Ser Gly Gly Asn Lys Arg Lys Leu Ser Thr Ala

980 985 990

Ile Ala Leu Ile Gly Cys Pro Pro Leu Val Leu Leu Asp Glu Pro Thr

995 1000 1005

Thr Gly Met Asp Pro Gln Ala Arg Arg Met Leu Trp Asn Val Ile Val

1010 1015 1020

Ser Ile Ile Arg Glu Gly Arg Ala Val Val Leu Thr Ser His Ser Met

1025 1030 1035 1040

Glu Glu Cys Glu Ala Leu Cys Thr Arg Leu Ala Ile Met Val Lys Gly

1045 1050 1055

Ala Phe Arg Cys Met Gly Thr Ile Gln His Leu Lys Ser Lys Phe Gly

1060 1065 1070

Asp Gly Tyr Ile Val Thr Met Lys Ile Lys Ser Pro Lys Asp Asp Leu

1075 1080 1085

Leu Pro Asp Leu Asn Pro Val Glu Gln Phe Phe Gln Gly Asn Phe Pro

1090 1095 1100

Gly Ser Val Gln Arg Glu Arg His Tyr Asn Met Leu Gln Phe Gln Val

1105 1110 1115 1120

Ser Ser Ser Ser Leu Ala Arg Ile Phe Gln Leu Leu Leu Ser His Lys

1125 1130 1135

Asp Ser Leu Leu Ile Glu Glu Tyr Ser Val Thr Gln Thr Thr Leu Asp

1140 1145 1150

Gln Val Phe Val Asn Phe Ala Lys Gln Gln Thr Glu Ser His Asp Leu

1155 1160 1165

Pro Leu His Pro Arg Ala Ala Gly Ala Ser Arg Gln Ala Gln Asp

1170 1175 1180

<210> 80

<211> 13

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> DD4

<220>

<221> 变体

<222> 12

<223> X= 任意氨基酸，优选Gly

<400> 80

Arg Asp Ser Phe Gly Ser Pro Pro Pro Met Ala Xaa Gly

1 5 10

<210> 81

<211> 4

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> DD5

<400> 81

Gly Phe Trp Ile

1

<210> 82

<211> 4

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> DD6

<400> 82

Arg Phe Lys Gly

1

<210> 83

<211> 4

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> DD7

<400> 83

Lys Phe Tyr Lys

1

<210> 84

<211> 8

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> 六聚组氨酸

<400> 84

Gly His His His His His His Gly

1 5

<210> 85

<211> 23

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> V12

<400> 85

Thr Asn Gly Ser Val Leu Arg Glu Phe Thr Leu Leu Glu Leu Glu Val

1 5 10 15

Val Thr Arg Asn Thr Glu Leu

20

<210> 86

<211> 21

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> T1

<400> 86

Ser Val Leu Glu Glu Asn Arg Pro Phe Ala Gln Gln Leu Ser Asn Val

1 5 10 15

Tyr Phe Thr Ile Leu

20

<210> 87

<211> 1176

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4N-1177

<400> 87

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly

1140 1145 1150

Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser

1155 1160 1165

Gln Arg Lys Gly Ser Glu Gly Thr

1170 1175

<210> 88

<211> 1097

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4C-1177

<400> 88

Cys Ser Cys Ser Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val

1 5 10 15

Asp Asp Leu Thr Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu

20 25 30

Met Asp Val Val Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys

35 40 45

Ile Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His

50 55 60

Arg Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp

65 70 75 80

Leu Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile

85 90 95

Phe Leu Lys Val Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly

100 105 110

Gly Ala Gln Gln Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu

115 120 125

Gly Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys

130 135 140

Ser Pro Gly Ala Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu

145 150 155 160

Pro Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu

165 170 175

Gln His Val Gln Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg

180 185 190

Ser His Lys Asp Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val

195 200 205

Phe Leu Ala Leu Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr

210 215 220

Pro Ala Leu Thr Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe

225 230 235 240

Phe Ser Met Asp Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp

245 250 255

Val Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly

260 265 270

Trp Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro

275 280 285

Ser Val Ser Pro Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr

290 295 300

Gln Val Asn Pro Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu

305 310 315 320

Thr Met Leu Pro Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro

325 330 335

Gln Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg

340 345 350

Asn Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser

355 360 365

Ser Leu Lys Ser Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile

370 375 380

Ser Ile Gly Gly Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu

385 390 395 400

Val Gly Phe Leu Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly

405 410 415

Pro Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His

420 425 430

Leu Glu Thr Glu Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp

435 440 445

His Ala Leu Val Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg

450 455 460

Ala Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val

465 470 475 480

Ile Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr

485 490 495

Val Leu Thr Thr Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe

500 505 510

Ser Met Ser Phe Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu

515 520 525

Arg Val Asn Lys Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro

530 535 540

Thr Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser

545 550 555 560

Val Ser Ala Gly Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys

565 570 575

Ala Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu

580 585 590

Leu Tyr Gly Trp Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu

595 600 605

Phe Asp Val Pro Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu

610 615 620

Phe Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe

625 630 635 640

Glu Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu

645 650 655

Leu Ile Val Phe Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu

660 665 670

Ala Leu Ser Gln Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu

675 680 685

His Ser Ala Asn Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe

690 695 700

Ala Met Val Val Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val

705 710 715 720

Gln Arg His Phe Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu

725 730 735

Pro Ile Val Asp Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile

740 745 750

Ile Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr

755 760 765

Lys Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val

770 775 780

Gly Val Arg Pro Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala

785 790 795 800

Gly Lys Thr Thr Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr

805 810 815

Ser Gly Asp Ala Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser

820 825 830

Glu Val His Gln Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp

835 840 845

Glu Leu Leu Thr Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg

850 855 860

Gly Val Pro Ala Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys

865 870 875 880

Ser Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser

885 890 895

Gly Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys

900 905 910

Pro Pro Leu Val Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln

915 920 925

Ala Arg Arg Met Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly

930 935 940

Arg Ala Val Val Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu

945 950 955 960

Cys Thr Arg Leu Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly

965 970 975

Thr Ile Gln His Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr

980 985 990

Met Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro

995 1000 1005

Val Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu

1010 1015 1020

Arg His Tyr Asn Met Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala

1025 1030 1035 1040

Arg Ile Phe Gln Leu Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu

1045 1050 1055

Glu Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe

1060 1065 1070

Ala Lys Gln Gln Thr Glu Ser His Asp Leu Pro Leu His Pro Arg Ala

1075 1080 1085

Ala Gly Ala Ser Arg Gln Ala Gln Asp

1090 1095

<210> 89

<211> 1178

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4N-1179

<400> 89

Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr

1 5 10 15

Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro

20 25 30

Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu

35 40 45

Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala

50 55 60

Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro

65 70 75 80

Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn

85 90 95

Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu

100 105 110

Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu

115 120 125

Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu

130 135 140

Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu

145 150 155 160

Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser

165 170 175

Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala

180 185 190

His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala

195 200 205

Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr

210 215 220

Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile

225 230 235 240

Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val

245 250 255

Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser

260 265 270

Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile

275 280 285

His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met

290 295 300

Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser

305 310 315 320

Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser

325 330 335

Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp

340 345 350

Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser

355 360 365

Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys

370 375 380

Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr

385 390 395 400

Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser

405 410 415

Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu

420 425 430

Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met

435 440 445

Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu

450 455 460

Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn

465 470 475 480

Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn

485 490 495

Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu

500 505 510

Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr

515 520 525

Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu

530 535 540

Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr

545 550 555 560

Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp

565 570 575

Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly

580 585 590

Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe

595 600 605

Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val

610 615 620

Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro

625 630 635 640

Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro

645 650 655

Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys

660 665 670

Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn

675 680 685

Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser

690 695 700

Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met

705 710 715 720

His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe

725 730 735

Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser

740 745 750

Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile

755 760 765

Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp

770 775 780

Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val

785 790 795 800

Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly

805 810 815

Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp

820 825 830

Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala

835 840 845

Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp

850 855 860

Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp

865 870 875 880

Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys

885 890 895

Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly

900 905 910

Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly

915 920 925

Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro

930 935 940

Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala

945 950 955 960

Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu

965 970 975

Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg

980 985 990

Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys

995 1000 1005

Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met

1010 1015 1020

Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu

1025 1030 1035 1040

Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn

1045 1050 1055

Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val

1060 1065 1070

Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro

1075 1080 1085

Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu

1090 1095 1100

Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met

1105 1110 1115 1120

Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly

1125 1130 1135

Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Phe Gly

1140 1145 1150

Thr Gly Leu Tyr Leu Thr Leu Val Arg Lys Met Lys Asn Ile Gln Ser

1155 1160 1165

Gln Arg Lys Gly Ser Glu Gly Thr Cys Ser

1170 1175

<210> 90

<211> 1095

<212> PRT

<213> 人工序列（Artificial Sequence）

<220>

<223> ABCA4C-1179

<400> 90

Cys Ser Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp Asp

1 5 10 15

Leu Thr Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met Asp

20 25 30

Val Val Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile Gly

35 40 45

Gln Glu Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg Ala

50 55 60

Tyr Ala Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu Gly

65 70 75 80

Leu Ser Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe Leu

85 90 95

Lys Val Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly Ala

100 105 110

Gln Gln Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly Pro

115 120 125

Arg Glu Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser Pro

130 135 140

Gly Ala Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro Glu

145 150 155 160

Cys Pro Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln His

165 170 175

Val Gln Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser His

180 185 190

Lys Asp Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe Leu

195 200 205

Ala Leu Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro Ala

210 215 220

Leu Thr Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe Ser

225 230 235 240

Met Asp Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val Leu

245 250 255

Leu Asn Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp Leu

260 265 270

Pro Glu Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser Val

275 280 285

Ser Pro Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln Val

290 295 300

Asn Pro Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr Met

305 310 315 320

Leu Pro Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln Arg

325 330 335

Thr Gln Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn Ile

340 345 350

Ser Asp Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser Leu

355 360 365

Lys Ser Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser Ile

370 375 380

Gly Gly Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val Gly

385 390 395 400

Phe Leu Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro Ile

405 410 415

Thr Arg Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu Glu

420 425 430

Thr Glu Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His Ala

435 440 445

Leu Val Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala Ser

450 455 460

Leu Pro Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile Ser

465 470 475 480

Gln Pro Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val Leu

485 490 495

Thr Thr Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser Met

500 505 510

Ser Phe Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg Val

515 520 525

Asn Lys Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr Thr

530 535 540

Tyr Trp Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val Ser

545 550 555 560

Ala Gly Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala Tyr

565 570 575

Thr Ser Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu Tyr

580 585 590

Gly Trp Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe Asp

595 600 605

Val Pro Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe Ile

610 615 620

Gly Ile Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu Asn

625 630 635 640

Asn Arg Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu Ile

645 650 655

Val Phe Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala Leu

660 665 670

Ser Gln Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His Ser

675 680 685

Ala Asn Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala Met

690 695 700

Val Val Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln Arg

705 710 715 720

His Phe Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro Ile

725 730 735

Val Asp Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile Thr

740 745 750

Gly Gly Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys Ile

755 760 765

Tyr Pro Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly Val

770 775 780

Arg Pro Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly Lys

785 790 795 800

Thr Thr Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser Gly

805 810 815

Asp Ala Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu Val

820 825 830

His Gln Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu Leu

835 840 845

Leu Thr Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly Val

850 855 860

Pro Ala Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser Leu

865 870 875 880

Gly Leu Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly Gly

885 890 895

Asn Lys Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro Pro

900 905 910

Leu Val Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala Arg

915 920 925

Arg Met Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg Ala

930 935 940

Val Val Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys Thr

945 950 955 960

Arg Leu Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr Ile

965 970 975

Gln His Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met Lys

980 985 990

Ile Lys Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val Glu