在人胚胎干细胞中高效表达PELO蛋白  

The Overexpression of PELO in Human Embryonic Stem Cells

作　　者：胡旭林[1] 纪家葵 

机构地区：[1]清华大学医学院干细胞与再生医学中心,北京100084

出　　处：《生物技术通报》2013年第10期170-176,共7页Biotechnology Bulletin

基　　金：清华大学自主科研计划(2010THZ03)

摘　　要：以PELO全长cDNA为模板,采用TOPO克隆的方法,将人的PELO基因克隆到pENTR-D-TOPO载体上,形成pENTR-D-PELO克隆载体。然后通过同源重组将含有EF1α启动子的pENTR-5’-EF1α和pENTR-D-PELO重组到表达载体p2k7上,形成EF1α-PELO-p2k7表达载体。将构建好的PELO高表达载体在293FT细胞中制备成相应的慢病毒表达载体,并用慢病毒侵染人胚胎干细胞,最终成功实现在人胚胎干细胞中稳定且高效地表达PELO蛋白。Using full length human PELO cDNA as template, TOPO cloned PELO gene to pENTR-D-TOPO vector to form the pENTR-D-PELO vector, then performed the MultiSite Gateway LR recombination reaction between pENTR-5’-EF1α and pENTR-D-PELO to get the EF1α-PELO-p2k7 expression vector. PELO overexpression lentivirus was produced in 293FT cells and ransducing human embryonic stem cells to overexpress PELO.

关 键 词：PELO 高表达 胚胎干细胞 慢病毒 同源重组 

分 类 号：Q78[生物学—分子生物学]