Stella基因真核表达载体的构建及其对小鼠胚胎干细胞多能性的影响  

Construction of Stella Eukaryotic Expression Vector and Its Effect on Pluripotency of Mouse Embryonic Stem Cells

作　　者：白耀富[1] 刘维帅 华进联[1] 

机构地区：[1]西北农林科技大学动物医学院、陕西省干细胞工程技术中心、农业部动物生物技术重点实验室,陕西杨凌712100 [2]杨棱示范区医院病理科,陕西杨凌712100

出　　处：《四川动物》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

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.

关 键 词：胚胎干细胞 减数分裂 多能性 

分 类 号：Q782[生物学—分子生物学] Q786