绵羊Sox2基因逆转录病毒载体的构建与检测

为建立利用内源诱导因子诱导绵羊体细胞为多能干细胞(induced pluripotent stem cell,iPS)的方法,对绵羊Sox2基因进行克隆,并与pMXs连接构建逆转录病毒载体,将构建的载体转染293GP细胞以获得假病毒上清,利用假病毒上清侵染绵羊胎儿成纤维细胞以检测细胞中的Sox2表达变化。PCR和酶切鉴定结果显示,成功构建了pMXs-Sox2重组质粒,该质粒具有转染293GP细胞的能力,所获得的假病毒上清侵染绵羊胎儿成纤维细胞后,可诱导细胞表达Sox2基因。本研究为开展绵羊iPS的相关研究提供依据。

Construction and Detection of Retrovirus Vector with Ovine Sox2 Gene

In order to construct the retroviral vector with sheep Sox2 to induce ovine somatic cell to iPS,sheep Sox2 gene was cloned,and then linked with retroviral vector pMXs.293GP cells were co-transfected by the vector and pVSV-G.Viral supernatant was infected sheep fibroblast cells in order to detect the expression of Sox2.The results were showed as followed.PCR and restriction enzyme digestion revealed that pMXs-Sox2 plasmid was constructed successfully.Expression of Sox2 was detected in sheep fibroblasts cell after being infected by the viral supernatant obtained.The results indicated that the vector obtained could be used to induce sheep somatic cell reprogramming.