慢病毒介导Nanog基因在小鼠ES细胞的表达  

Expression of Nanog Gene with the Mediation of Ientiviral Vector in Mouse ES Cells

作　　者：曹鸿国[1] 殷慧群[1] 张卫琴[1] 陈涛[1] 黄伟玲[1] 

机构地区：[1]安徽农业大学动物科技学院,合肥230036

出　　处：《中国生物工程杂志》2009年第5期23-27,共5页China Biotechnology

摘　　要：试验尝试构建小鼠Nanog基因慢病毒表达载体,培养表达外源Nanog基因的小鼠ES细胞。结果显示通过RT-PCR扩增出918bp的小鼠Nanog基因,测序正确的小鼠Nanog基因通过慢病毒介导在小鼠ES细胞表达后,表达外源Nanog基因的小鼠ES细胞生长状态同普通ES细胞无明显差异,在无LIF的ES细胞培养液培养条件下,表达外源Nanog基因的小鼠ES细胞保持正常的ES细胞集落,碱性磷酸酶、Oct4和SSEA-1免疫细胞化学检测为阳性,相同情况下未表达外源Nanog基因的小鼠ES细胞集落退化消失。试验证实了通过慢病毒载体介导培养了表达外源Nanog基因的小鼠ES细胞。In order to further study mouse embryonic stem cells （ES cells）, lentiviral vector PLL-IRES- Nanog-Neo was constructed. Mouse ES cells overexpressed nanog by mediation of lentiviral were cultured on mouse fetal fibroblast feeders after 2 weeks under G418 media and examined according to gowth characteristics. Results were showed that 918 bp nanog fragments were expressed in mouse ES cells mediated by lentiviral vector PLL-IRES-Nanog-Neo, mouse nanog-ES cells were taken on mass-like image and positve with alkaline phosphatase staining and Oct4 and SSEAI immunocytochemistry under no LIF condition in the media. It is concluded that mouse ES cells Elevated nanog gene expression by mediation of lentiviral were constucted and cultured.

关 键 词：小鼠 ES细胞 NANOG基因 表达 

分 类 号：Q811.4[生物学—生物工程] Q78