中国现代医学杂志2017，Vol.27Issue(26)：25-29,5.DOI:10.3969/j.issn.1005-8982.2017.26.005
HCV NS4B表达载体的构建及其对细胞凋亡的影响
Construction of expression vector of HCV NS4B and its effect on cell apoptosis
摘要
Abstract
Objective To construct an eukaryotic expression vector of Hepatitis C virus (HCV) nonstructural protein NS4B,and investigate the effect of this protein on apoptosis of U2OS cells.Methods The fragment of target gene NS4B was obtained by PCR,and connected with the PCMV-tag2b vector using the homologous recombination method,and the PCMV-tag2b-NS4B was transformed into the E.coli DH5a.The right recombinant plasmid was selected.Lipofectamine 2000 was used to transfect PCMV-tag2b-NS4B into U2OS cells.Western blot was applied to detect the expression of HCV NS4B protein in U2OS cells,and immunofluorescence (IF) was used to assess the apoptosis of U2OS cells.Results The data showed that the recombinant plasmid PCMV-tag2b-NS4B was consistent with the one published on NCBI and could express HCV NS4B protein in U2OS cells.DAPI staining showed that the apoptosis rate of the PCMV-tag2b-NS4B group was (17.25 ± 2.95)％ and that of the control group was (6.53 ± 2.36)％.Conclusions The eukaryotic expression vector of HCV NS4B has been successfully constructed,and HCV NS4B protein is expressed in U2OS cells and induces apoptosis.关键词
丙型肝炎病毒/非结构蛋白4B/重组质粒/凋亡Key words
Hepatitis C virus/nonstructural protein 4B/recombinant plasmid/apoptosis分类
医药卫生引用本文复制引用
宋彬妤,赵嘉伟,宋静,陈思思,胡志廷,韩昕,王敏敏,任来峰..HCV NS4B表达载体的构建及其对细胞凋亡的影响[J].中国现代医学杂志,2017,27(26):25-29,5.基金项目
山西省自然科学基金(No:2014021037-9) （No:2014021037-9）
山西省高等学校大学生创新创业训练项目(No:2014052) （No:2014052）
山西医科大学汾阳学院科技发展基金(No:2016B02) （No:2016B02）
