摘要
为了建立体细胞重编程过程中检测Sox2基因表达变化的方法,本研究利用克隆获得的小鼠Sox2基因与逆转录病毒载体pMX-IRES-GFP连接,构建带有报告基因GFP的逆转录病毒载体,将其转染293GP细胞后获得假病毒上清。将获得假病毒上清侵染小鼠成纤维细胞(NIH3 T3细胞)后观察GFP的表达和检测Sox2转录量的变化。其结果,成功构建带有GFP的Sox2基因逆转录病毒载体pMX-Sox2-IRES-GFP;将构建的载体转染后293细胞后获得能侵染NIH3T3细胞的假病毒上清,且与未侵染的NIH3 T3细胞相比,侵染后的NIH3T3细胞的Sox2表达量提高近10倍。本研究为开展在体细胞重编程过程中Sox2表达与重编程效率的相关性提供依据。
In order reprogrammlng, to establish the method detecting of Sox2 this study cloned mouse Sox2 gene, and gene expression during somatic cells linked with retroviral vector pMX- IRES-GFP. 293GP cells were transfected with the vector and pVSV-G vector, mouse fibroblasts （NIH3T3 cells） were infected by viral supematant. GFP expression was observed and the amount of Sox2 transcription was tested after infection. The results were showed as followed. The retroviral vector pMX-Sox2-IRES-GFP was constructed successfully, viral supematant was obtained after the vector and pVSV-G co-transfection. Compared to negative control infected NIH3T3 cells expression of Sox2 increased nearly 10-fold.These results would be used for promoting somatic cells reprogramming study.
出处
《黑龙江动物繁殖》
2012年第1期8-11,共4页
Heilongjiang journal of animal reproduction
基金
国家自然科学基金资助项目(31000990
30771538)
中央高校基本科研业务费专项资金资助(DL10BA06)
作者简介
渠俊杰（1985-），女，研究生。
通讯作者：朴善花，高级畜牧师。E-mail：antiezhu@tom．com
