MicroRNA122真核表达载体的构建及在HepG2细胞中的表达与检测  

Construction and Detection of Eukaryotic Expression Vector for MicroRNA122

作　　者：郝美君[1] 张祯祯[1] 张文露[1] 曾爱中[1] 崔静[1] 黄艺丹[1] 黄爱龙[1] 

机构地区：[1]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《生物技术通报》2008年第6期144-146,151,共4页Biotechnology Bulletin

基　　金：973计划前期研究专项(2007CB516810)

摘　　要：为进一步研究microRNA的功能及作用靶标提供技术平台,构建microRNA真核表达载体。从HepG2215细胞基因组DNA扩增microRNA122前体,克隆到pEGFP-C1的载体上,经测序鉴定后转染至HepG2细胞中,荧光显微镜观察绿色荧光蛋白表达情况并提取基因组总RNA,RT-PCR检测microRNA122表达,TA克隆鉴定。结果表明,成功构建microRNA122真核表达载体,并证实其在真核细胞HepG2中表达。此方法成功构建microRNA表达,并为后续研究提供良好的技术平台。The eukaryotic expression vector of PEGFP-c1-micro122 was constructed to research the function of mircroRNA122. The pre-mircroRNA122 was amplified from total DNA of hepatoblastama cell line HepG2215. Gene from HepG2215 was inserted into PEGFP-c1 vector to construct a vector PEGFP-c1- microRNA122 for fusion protein. The expression of the fusion protein in HepG2 cell transfect with the vector was evaluated by the expression of GFP. Total RNA was extracted and the expression of microRNA122 was evaluated by RT-PCR and sequencing. As a result, the recombinant PEGFP-c1- microRNA122 was successfully constructed in Pegfp-c1 plasmid and the expression of microRNA122 with EGFP was detected.

关 键 词：MICRORNA 绿色荧光蛋白 真核表达载体 

分 类 号：Q78[生物学—分子生物学]