| 盛陶 曾建平 李辉.ZO-1 基因的慢病毒载体构建[J].现代生物医学进展英文版,2014,14(4):628-630. |
| ZO-1 基因的慢病毒载体构建 |
| Construction of Lentiviral Vector Carrying ZO-1 Gene |
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| DOI： |
| 中文关键词: ZO-1 基因克隆 慢病毒载体 |
| 英文关键词: ZO-1 Gene cloning Lentiviral vector |
| 基金项目:湖南省科学技术厅科技计划重点项目（2012SK2015） |
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| 中文摘要: |
| 目的：相邻两心肌纤维的连接处称心肌闰盘，它在心肌的机电活动中扮演重要角色。目前有关闰盘相关基因Cx43 的研究已
被广泛关注，而在不同心肌病理状态下，其变化及功能都与另一闰盘相关基因ZO-1 有极为密切的联系。目前有关ZO-1 的研究比
较少，此次欲将猪ZO-1 基因克隆到慢病毒表达载体并进行鉴定，为进一步研究其功能奠定基础。方法：利用聚合酶链反应（PCR）
扩增猪ZO-1 基因，并克隆到慢病毒表达载体，通过转化、抽提质粒、双酶切和测序鉴定构建的Lenti-EF1α-EGFP-TRE-ZO-1 重组
载体。结果：猪ZO-1 基因片段重组到慢病毒载体；PCR和双酶切鉴定，电泳结果显示均能得到与理论大小相符的片段；经测序证
实成功构建Lenti-EF1α-EGFP-TRE-ZO-1 慢病毒表达载体。结论：通过一系列实验证实Lenti-EF1α-EGFP-TRE-ZO-1 慢病毒表达
载体构建成功，可利用其感染相关细胞或注入动物体内，为后续的体外及在体研究奠定基础，明确其在不同心肌病理状态下所发
挥的独特作用。 |
| 英文摘要: |
| Objective:The joint of adjacent heart muscle fibers is called myocardial intercalated disc, which is important for
myocardial electrical activity. It has been widely concerned about the study of intercalated disc related gene Cx43. Under different
pathological status, it is related to another gene ZO-1. However, the research of ZO-1 is scanty, so we cloned the ZO-1 gene of porcine
into the lentiviral vector and identificate the recombinant plasmid，which will lay the foundation for further research.Methods:The
porcine ZO-1 gene was amplified by PCR, and cloned into lentiviral expression vector. The reconstructed Lenti- EF1α-EGFP-TRE-ZO-1
plasmid was identified by transformation, plasmid extraction, double enzyme digestion and DNA sequencing analysis.Results:The
porcine ZO-1 DNA sequences were successfully inserted into the lentiviral vector.The reconstructed plasmid was identified correctly by
PCR and double enzyme digestion．The recombinant Lenti-EF1α-EGFP-TRE-ZO-1 vector was confirmed by DNA sequencing.Conclusion:Lenti-EF1α-EGFP-TRE-ZO-1 lentiviral vector has been successfully constructed, so we can use it to infect related cells or
animals, which will be useful for further study in vitro and in vivo, to understand its function in different pathological status. |
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