人microRNA-146a慢病毒表达载体的构建及鉴定  

Construction and function identification of recombinant lentivirus vector expressing has-miR-146a

作　　者：李景怡[1,2] 万瑛[1] 张晋宇[1] 邹丽云[1] 李娜[1] 刘婷[1] 柴琳琳[1] 熊锐华 吴玉章[1] 

机构地区：[1]第三军医大学全军免疫学研究所,重庆400038 [2]第三军医大学第一附属医院风湿病中心,重庆400038

出　　处：《免疫学杂志》2008年第6期671-673,共3页Immunological Journal

基　　金：国家自然科学基金(30801030)

摘　　要：目的构建针对has-miR-146a的慢病毒表达载体,并鉴定成熟has-miR-146a在细胞内表达水平。方法PCR扩增pri-miR-146a,克隆于慢病毒载体plenti-GFP中,转染293FT细胞,收获并浓缩慢病毒颗粒,感染Jurkat细胞。结果成功构建了has-miR-146a的慢病毒表达载体,建立了高效转染系统。结论建立了高效稳定表达has-miR-146a的慢病毒转染系统。Objective To construct a lentivirus vector expressing microRNA （miRNA） miR-146a and transfect the vector into Jurkat cell line. Methods PrimiR-146a amplified by PCR was inserted into plenti-GFP vector, and then identified by restriction endonuclease digestion and nucleotide sequencing. Jurkat cell line was transfected with the plasmid pluG-miR-146a. The expression of miR-146a was detected by fluorescence microscopy, flow cytometry, and Real-time PCR. Results The sequence of miR-146a was contained in the constructed recombinant plasmid. After transfection with the plasmid, miR-146a could be expressed in Jurkat cell line. Conclusion The constructed lentivirus vector can express miR-146a in vitro, which is the basis for further study on the function of miR-146a.

关 键 词：microRNA-146a 慢病毒表达载体 JURKAT细胞 

分 类 号：R346[医药卫生—基础医学]