郑州大学学报（医学版）2011，Vol.46Issue(1)：19-22,4.
小鼠微小RNA294逆转录病毒载体的构建及鉴定
Construction and identification of embryonic stem cell-specific microR-NA294 retroviral vector of mice
摘要
Abstract
Aim: To construct microRNA-294 ( miR-294 ) retroviral vector. Methods: pri-miR-294 amplified by PCR was inserted into pMX-IRES-GFP vector, and then identified by nucleotide sequencing. PLAT-E cells were transfected with retroviral vector pMX-miR-294-IRES-GFP. All virus stocks were produced by calcium phosphate-mediated transfection. The titer of virus was tested according to the expression level of CFP. Results: DNA sequencing demonstrated that pMX-miR-294-IRES-GFP was successfully constructed. The titer of concentrated virus was (1 ~ 5) × 106 TU/mL. Conclusion: pMX-miR-294-IRES-GFP retroviral vector was successfully constructed, which paved the way for the induction of induced pluri-potent stem cells and the investigation of cellular reprogramming mechanism.关键词
微小RNA/细胞重编程/逆转录病毒载体/小鼠Key words
microRNA/ cellular reprogramming/ retro viral vector/ mouse分类
生物科学引用本文复制引用
许杰,闫秋月,湛彦强,张苏明..小鼠微小RNA294逆转录病毒载体的构建及鉴定[J].郑州大学学报（医学版）,2011,46(1):19-22,4.基金项目
国家自然科学基金资助项目 30370505 （）
武汉市科技计划基金资助项目 200860423216 （）
