吉林大学学报（医学版）Issue(6)：1149-1154,6.DOI:10.13481/j.1671-587x.20140605
携带FLAG标签的人BTG2真核表达载体的构建及其在HeLa细胞中的表达
Construction of human BTG2 eukaryotic expression vector with FLAG tag and its expression in HeLa cells
摘要
Abstract
Objective To construct an eukaryotic expression vector of human B-cell translocation gene 2 (BTG2), to express the FLAG-tagged BTG2 protein in HeLa cells,and to supply an experimental tool for investigating the function of BTG2 gene.Methods The full-length BTG2 fragment was obtained by PCR and inserted into the multiple cloning site of pcDNA3.1 (+)vector. Oligo DNA encoding FLAG tag was designed and inserted into pcDNA3.1(+)-BTG2 to construct another vector pcDNA3.1(+)-FLAG-BTG2.The HeLa cells were divided into pcDNA3.1(+)empty vector group,pcDNA3.1(+)-BTG2 group and pcDNA3.1(+)-FLAG-BTG2 group.The HeLa cells were transfected with recombinant plasmids.Western blotting using anti-FLAG antibody was performed to detect the expression of FLAG-BTG2 protein in HeLa cells.Results The sequence of the vector was verified by both BamH Ⅰ endonuclese digestion and DNA sequencing. The Western blotting analysis confirmed that FLAG-fused BTG2 was detected in pcDNA3.1(+)-FLAG-BTG2 group but not in empty vector or pcDNA3.1(+)-BTG2 groups. Conclusion The eukaryotic expression vector pcDNA3.1(+)-FLAG-BTG2 is successfully constructed and FLAG-tagged BTG2 protein is expressed in HeLa cells.关键词
B细胞易位基因 2/抑癌基因/FLAG标签/真核表达载体/融合蛋白Key words
B-cell translocation gene 2/tumor suppressor gene/FLAG tag/eukaryotic expression vector/fusion protein分类
生物科学引用本文复制引用
赵金匣,王志平,陶燕,贺振华,郭琦,洪梅..携带FLAG标签的人BTG2真核表达载体的构建及其在HeLa细胞中的表达[J].吉林大学学报（医学版）,2014,(6):1149-1154,6.基金项目
国家自然科学基金青年基金资助课题（81302240）；甘肃省科技计划项目资助课题（1308RJYA056）；兰州大学中央高校基本科研业务费项目资助课题 （）
