抗凋亡基因bcl-2转染哺乳动物细胞的研究  

Study on the Transfection of the Apoptosis -Resistant Gene bcl-2 into Mammalian Cell

作　　者：何庆[1] 苏京[2] 刘铭[3] 王保平[2] 王晶[2] 陈克勤[2] 尹潍[1] 

机构地区：[1]天津医科大学总医院内分泌科,300052 [2]丹麦诺和诺德中国研究发展中心 [3]天津医科大学总医院老年病研究室,300052

出　　处：《天津医药》2006年第2期105-107,共3页Tianjin Medical Journal

基　　金：天津市自然科学基金资助课题(项目编号:983702411;043607911)

摘　　要：目的:构建抗凋亡基因bcl-2的哺乳动物细胞表达型质粒并转染哺乳动物细胞293T细胞。方法:利用基因重组技术,将bcl-2全编码cDNA序列,导入pCMV-Tag1表达型质粒;应用磷酸钙共沉淀技术,将其转染293T细胞;并利用免疫沉淀和Westernblotting技术检测Bcl-2融合蛋白的表达。结果:bcl-2全编码cDNA序列成功导入pCMV-Tag1表达型质粒;在转染后的293T细胞成功地检测出bcl-2融合蛋白的表达,融合蛋白大小与预期完全一致。结论:该表达型质粒可以成功转染哺乳动物细胞,为对胰岛细胞的转染奠定了基础。Objective： To construct the mammalian expression plasmid of bcl-2 gene, which is believed to have the effect of resisting the apoptosis, and transfect the mammalian cell with it. Methods： The full length cDNA of bcl-2 gene was put into expression plasmid pCMV-Tag 1 by gene recombination method. The 293T cell lines were transfected with the bc1-2 expression plasmid through the calcium phosphate coprecipitation technology. The expression of bcl-2 fusion protein was detected with the immunoprecipitation and Western blotting technology. Results： Full length cDNA of bc1-2 gene was constructed into the expression plasmid of pCMV-Tag 1 successfully. Bcl-2 fusion protein had been detected successfully in transfected 293T cell lines, and the size of the fusion protein was the same as expected. Conclusion： This study indicated that the expression plasmid can express in cultured mammalian cells and can be used to transfect the apoptosis-resistant gene bcl-2 into the islet cell.

关 键 词：基因 bcl-2质粒基因 DNA 重组 转染 免疫印迹法 

分 类 号：R392.2[医药卫生—免疫学] R394[医药卫生—基础医学]