RPS27a蛋白重组质粒的构建及其在肝癌细胞株的定位

目的:构建pcDNA3.1-RPS27a重组质粒载体并观察其在HepG2和SMMC7721细胞株中的定位和表达情况.方法:使用基因合成法合成RPS27a基因,将扩增后的目的基因连接至pcDNA3.1载体.转化DH5α大肠埃希菌后,进行质粒抽提然后电泳及测序鉴定.将鉴定后的pCDNA3.1-RPS27a质粒分别转染HepG2和SMMC7721细胞株,通过激光共聚焦显微镜观察其在真核细胞中的表达和定位情况.结果:pCDNA3.1-RPS27a 重组质粒构建成功.通过激光共聚焦显微镜观察发现RPS27a主要表达于HepG2和SMMC7721细胞的胞质中.结论:pCDNA3.1-RPS27a载体构建成功,证实了RPS27a主要表达于HepG2和SMMC7721细胞的细胞质中.

Construction of RPS27a recombinant plasmid vector and its localization in hepatoma cell lines

Objective:To construct the recombinant plasmid vector of pcDNA3.1-RPS27a,and investigate the localization and expression of RPS27a protein in HepG2 and SMMC7721 cell lines.Methods:The RPS27a gene was synthesed and cloned into the pcDNA3.1 vector.After the pcDNA3.1 plasmid was transfected into E.coli DH5α,and the positive plasmid was extracted and sequenced.The reconstructed pcDNA3.1-RPS27a plasmid was transfected into the HepG2 and SMMC7721 cell lines.The expression and localization of RPS27a in eukaryotic cells were detected by laser confocal microscopy.Results:The recombinant plasmid pcDNA3.1-RPS27a was successfully constructed.The laser confocal microscopy showed that the RPS27a mainly expressed in the cytoplasm of HepG2 and SMMC7721 cell lines.Conclusions:The recombinant plasmid pcDNA3.1-RPS27a is successfully constructed,and the RPS27a protein mainly expresses in the cytoplasm of HepG2 and SMMC7721 cell lines.