吉林大学学报（医学版）2017，Vol.43Issue(4)：694-697,前插1,5.DOI:10.13481/j.1671-587x.20170406
miR-137过表达慢病毒的包装和鉴定
Packaging and identification of miR-137 overexpression lentivirus
摘要
Abstract
Objective:To construct lentiviral vector which can overexpression miR-137 and produce lentivirus by lentivirus packaging system, and to explore its infection efficiency and expression in HEK293T cells.Methods: miR-137 sequence was chemically synthesized and cloned into lentiviral vector GV209, and the recombinant plasmid containing human miR-137 was obtained and identified.Then miR-137 recombinant plasmid together with two helper plasmids were transfected into HEK293T cells using Lipofectamine 2000.After the HEK293T cells were infected in multiplicity of infection(MOI) 40 for 48 h, the expression of green fluorescent protein (GFP) was observed by fluorescence microscope and the expression level of miR-137 was detected by fluorescence quantitative PCR.Results:The sequencing results showed that the inserted gene sequence was completely consistent with the published human miR-137 gene sequence in GenBank.The GFP was observed in the HEK293T cells infected with miR-137 overexpression lentivirus under fluorescence microscope.The fluorescence quantitative PCR results showed that the expression level of miR-137 in the cells infected with overexpression lentivirus was 12.74 times higher than that in the control cells.Conclusion:The lentivirus containing miR-137 gene is successful packaged, and it could efficiently infect the HEK293T cells.关键词
miR-137/慢病毒/HEK293T细胞/绿色荧光蛋白Key words
miR-137/lentivirus/HEK293T cell/green fluorescent protein分类
生物科学引用本文复制引用
律东,梁春梅,李明颖,殷静雯,罗旭东,林举达,马国达..miR-137过表达慢病毒的包装和鉴定[J].吉林大学学报（医学版）,2017,43(4):694-697,前插1,5.基金项目
国家自然科学基金资助课题(81670252) （81670252）
广东省科技厅自然科学基金资助课题(2015A030313523) （2015A030313523）
广东省湛江市科技局科技计划项目资助课题(2016A01008) （2016A01008）
