摘要
构建Stella基因真核表达质粒,转染小鼠胚胎干细胞(Embryonic stem cells,ESC)并初步探讨Stella对减数分裂起始相关基因(Stra8)及胚胎干细胞多能性的影响。通过RT-PCR扩增目的基因,并连接至真核表达载体pEGFP-C1,利用重组质粒转染小鼠胚胎干细胞。对转染细胞进行荧光检测,确认Stella的表达,并利用免疫荧光及PCR检测转染细胞基因表达情况。酶切鉴定及测序分析表明成功构建含Stella基因的重组真核表达质粒,过表达Stella对ES细胞的增殖和形态学特征、进入减数分裂阶段的相关基因及其多能性基因的表达影响并不显著。故此得出结论:Stella在小鼠胚胎干细胞中能够正确表达,但对ES细胞的分化、Stra8基因的表达及其多能性基因的表达并无显著影响。
To explore the effects of recombinant vector Stella on the pluripotency of mouse embryonic stem cells （ESCs） and expression profile of specific meiosis marker-Stra8 in transfected ESCs. The eukaryotic expression vector containing Stel- la was constructed and transfected into ESCs. Expression profiles of GFP was observed in transfected ESCs, and expression levels of pluripotent genes and specific meiosis markers were simuhaneously detected by RT-PCR and immunofluorescent staining. The results demonstrated that although Stella which had been cloned in the recombinant vector was successfully ex- pressed in ESCs as identified by GFP and PCR amplification, it had no significant effect on the expression profiles of pluri- potent and specific meiosis marker genes.
出处
《四川动物》
CSCD
北大核心
2014年第1期118-122,共5页
Sichuan Journal of Zoology
基金
教育部博士点基金(RFDP,20120204110030)
国家自然科学基金(No.31272518)资助
关键词
胚胎干细胞
减数分裂
多能性
embryonic stem cells
meiosis
pluripotency
