人miR-106b的克隆及其慢病毒表达载体构建  

Cloning of hsa-miR-106b and construction of its lentiviral expression vector

作　　者：郑林[1] 王爽[1] 丁彦青[1] 

机构地区：[1]南方医科大学南方医院病理科,广东广州510515

出　　处：《中国热带医学》2011年第6期651-653,共3页China Tropical Medicine

基　　金：国家自然科学基金(81071735);国家自然科学青年基金(81000953);广东省高校科技创新重点项目(cxzd1016);广东省自然科学基金博士启动项目(10451051501004710)

摘　　要：目的克隆微小RNA hsa-miR-106b并构建其慢病毒表达载体。方法将PCR扩增得到的miR-106b前体序列和pLVTHM载体经双酶切后连接,产生pLVTHM-miR-106b慢病毒表达载体,双酶切后测序鉴定,筛选阳性克隆。用pLVTHM-miR-106b、psPAX2和pMD2.G质粒共转染包装细胞293FT,包装产生慢病毒。结果经双酶切鉴定和测序证实,成功构了miR-106b的慢病毒表达载体pLVTHM-miR-106b。倒置荧光显微镜下观察可见包装细胞293FT呈绿色荧光。结论成功构建了has-miR-106b的慢病毒表达载体,为深入研究miR-106b的生物学功能奠定基础。Aim To construct a lentiviral expression vector for hsa-miR-106b. Methods The pre-mir-106b amplified by PCR was inserted into pLVTHM. The recombinant plasmid pLVTHM-miR-106b was confirmed by restriction endonuclease analysis and DNA sequence. 293FT cells were cotransfected with lentivirus vector pLVTHM-miR-106b, psPAX2 and pMD2.G. All virus stocks were produced by calcium phosphate-mediated transfection. Results Restriction enzyme digestion and DNA sequencing demonstrated that the lentivirus vector pLVTHM-miR-106b was constructed successfully, Conclusion The successfully construction of lentivirus vector pLVTHM-miR-106b provides the basis for the further study of moleeular function of mir-106b.

关 键 词：hsa—miR-106b 肿瘤 慢病毒 

分 类 号：R739.6[医药卫生—肿瘤]