RPS27a蛋白重组质粒的构建及其在肝癌细胞株的定位

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

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。

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.