@article{YKXB3654,
author = {李 春梅 和 卓 业鸿 和 陈 梦飞 和 孙 雪荣 和 祁 影 和 葛 坚},
title = {原发性开角型青光眼致病基因小鼠MYOC Pro356Leu突变型质粒构建及鉴定},
journal = {眼科学报},
volume = {25},
number = {1},
year = {2016},
keywords = {},
abstract = {目的:构建小鼠MYOC Pro356Leu突变型质粒及鉴定。方法:1.将含有红重组酶基因的质粒pKD46电击入RP23-180F15 BAC克隆。2.利用PCR扩增pStart-K载体,将PCR产物电穿孔入RP23-180F15 BAC红色重组细菌的MYOC基因组片段。3.PCR扩增mMyoc AfeⅠ片段,亚克隆mMyoc AfeⅠ片段进入pBluescript载体。4.设计两条寡核苷酸将Pro356Leu(C to T at codon 356)导入pBS_mMyoc AfeⅠ克隆。5.通过酶切及连接酶反应将pBS_mMyoc AfeⅠmut clone导入pStart-K_mMyoc构建成pStart-K_mMyoc mut克隆。结果:成功构建小鼠MYOC Pro356Leu突变型质粒pStart-K_mMyoc mut克隆,并经酶切及测序证实。结论:应用红重组技术及pStart-K载体克隆大片段DNA是有效的DNA克隆方法,我们所构建的MYOC Pro356Leu突变型质粒将为研究MYOC基因在原发性开角型青光眼发病中的功能和作用提供实验依据。},
url = {https://ykxb.amegroups.com/article/view/3654}
}