小鼠microRNA302a逆转录病毒载体的构建及鉴定  

Construction and Identification of a Retroviral Vector Expressing MicroRNA302a

作　　者：许杰[1] 闫秋月[1] 湛彦强[1] 张苏明[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经科,武汉430030

出　　处：《神经损伤与功能重建》2010年第1期2-4,共3页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金资助项目(No.30370505);武汉科技计划项目(No.200860423216)

摘　　要：目的:为进一步研究microRNA在细胞重编程机制中的作用,构建microRNA302a(miR-302a)逆转录病毒载体。方法:从小鼠基因组DNA扩增pri-miR302a,克隆至pMx-IRES-GFP逆转录病毒载体上,测序鉴定,转染PLAT-E细胞包装逆转录病毒颗粒,并进行病毒滴度测定。结果:成功构建表达miR-302a的逆转录病毒载体pMx-miR302a-IRES-GFP,阳性克隆测序结果与目标基因序列完全一致。结论:pMx-miRNA302a-IRES-GFP载体构建成功。Objective： To construct a retroviral vector expressing miR-302a to investigate the role of micro- RNA in cellular reprogramming. Methods.. miR-302a precursor sequence was amplified from the mice genomic DNA, inserted into pMX-IRES-GFP vector and identified by DNA sequencing. PLAT-E cells were transfected with the pMx-miR302a-IRES-GFP and the titer of virus was tested by the expression of GFP. Results： It was demonstrated by DNA sequencing that the retroviral vector pMx-miR302a-IRES-GFP was successfully constructed. The titer of concentrated virus was （1 - 5） × 10^6 TU/mL. Conclusion： The retroviral vector pMx-miR302a-IRES-GFP was successfully constructed.

关 键 词：miRNA302a 逆转录病毒载体 细胞重编程 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学]