支原体污染对重组逆转录病毒的产生和感染靶细胞的影响  

Influence of Mycoplasma contamination on production of recombinant retrovirus and infection of target cells

作　　者：周佽想[1,2] 郭萌[1] 李稻[1] 赵倩[1,2] 

机构地区：[1]上海交通大学基础医学院病理生理学教研室,上海200025 [2]中国科学院上海生命科学研究院/上海交通大学医学院健康科学研究所

出　　处：《上海交通大学学报（医学版）》2008年第12期1527-1530,共4页Journal of Shanghai Jiao tong University:Medical Science

基　　金：上海市科委基金(06JC14074);国家高技术研究发展计划("863"计划)(2007AA02Z318)~~

摘　　要：目的探讨细胞支原体污染对重组逆转录病毒的产生和靶细胞感染效率的影响。方法利用倒置荧光显微镜或流式细胞仪检测绿色荧光蛋白(GFP)表达,确定细胞转染效率及病毒感染效率。PCR方法扩增支原体特异性核酸片段以检测培养细胞293T、NIH3T3及32D有无支原体污染。结果支原体抗生素Plasmocin有效清除了培养细胞内及上清中所污染的支原体;通过对细胞支原体的清除明显提高了逆转录病毒重组表达质粒在包装细胞中的转染效率及病毒感染靶细胞的效率。结论细胞受支原体污染后不仅影响了逆转录病毒重组表达质粒在包装细胞中的转染,而且严重影响了重组逆转录病毒对靶细胞的感染效率;Plasmocin是清除细胞支原体污染的理想抗生素。Objective To investigate the influence of Mycoplasma on the production and infection of recombinant retrovirus on target cells. Methods The expression of green fluorescence protein （GFP） was detected using inverted fluorescence microscop or flow cytometer. The efficiency of transfection and infection was analyzed. PCR was employed to determine whether cultured cells 293T, NIH3T3 and 32D were contaminated by Mycoplasm. Results Antibiotic Plasmocin effectively eliminated the Mycoplasma contamination both inside the cells and in the cell culture medium. After elimination of the Mycoplasma in the host cells with Plasmocin, both the production of the recombinant retrovirus and the infection efficiency on target cells were greatly improved. Conclusion Mycoplasma contamination can affect not only the production of retrovirus but also the infection of target cells. .Plasmocin is an ideal antibiotic for Mycoplasma elimination in the cultured cells.

关 键 词：支原体 病毒载体 基因转染 转染效率 

分 类 号：R346[医药卫生—基础医学]