pEGFP-N1-PGC-1α重组质粒的构建及体外表达  

Construction and Expression of pEGFP-N1-PGC-1α Recombinant Plasmid in Vitro

作　　者：于波海[1,2] 范世珍 张萌[2] 柴淼[2] 苏丽菊[2] 郑力辉[2] 韩笑[2] 毕佳冰 腾旭[1] 

机构地区：[1]哈尔滨医科大学生物学博士后流动站,黑龙江哈尔滨150081 [2]哈尔滨市第一医院检验科,黑龙江哈尔滨150010 [3]深圳市福田区中医院检验科,广东深圳518034

出　　处：《标记免疫分析与临床》2016年第7期820-824,共5页Labeled Immunoassays and Clinical Medicine

基　　金：黑龙江省卫生计生委科研课题No.2014-004;深圳市科创委研究项目JCYJ20150331091010278

摘　　要：目的构建PGC—1α真核表达载体pEGFP—N1-PGC-1α重组质粒,鉴定及体外表达。方法通过RT-PCR方法从组织中克隆PGC-1α基因片段,酶切后将其定向插入真核表达载体pEGFP—N1中,构建pEGFP—N1-PGC-1α重组质粒。结果双酶切法、测序法鉴定表明目的基因正确插入质粒。结论成功构建了pEGFP—N1-PGC-1α真核表达载体,转染huh-7细胞后,可瞬时、有效表达,为进一步研究PGC-1α调节HBV、HPV生物合成奠定了基础。Objective To construct eukaryotic expression plasmid carrying PGC-1α gene, and observe its expression in huh-7 cells. Methods PGC-1α gene was obtained from tissue through RT-PCR method and inserted eukaryotic expression vector pEGFP-N1. Thus pEGFP-N1-PGC-1α recombinant plasmid was constructed and transfected into huh-7 cells. Results The inserted fragment was identified by double enzyme digestion and DNA sequencing. Conclusion pEGFP-N1-PGC-1α recombinant plasmid was constructed and expressed successfully in huh-7 cells, which lays the foundation for future studies on the regulation of PGC-1α on HBV and HPV biosynthesis.

关 键 词：PGC-1Α pEGFP-N1真核表达载体 HUH-7细胞 

分 类 号：Q78[生物学—分子生物学] R346[医药卫生—基础医学]