瘦素基因在人胎盘间充质干细胞中转染与表达  

Transfection and expression of leptin gene in human placental mesenchymal stem cells

作　　者：朱振威[1] 刘春丽[2] 王博蔚[3] 陈一夫[1] 王占义[4] 高贝[1] 刘志辉[1] 

机构地区：[1]吉林大学口腔医院口腔修复科,长春130021 [2]吉林大学口腔医院口腔颌面外科,长春130021 [3]吉林大学第二医院妇产科 [4]长春市口腔医院口腔颌面外科

出　　处：《实用口腔医学杂志》2014年第4期487-491,共5页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:30471804);吉林省自然科学基金(编号:201015201);吉林大学"大学生创新性实验计划"项目(编号:2012A78211)

摘　　要：目的:构建瘦素(leptin)基因真核表达载体,将其转染至人胎盘间充质干细胞(HPMSCs)中并鉴定其表达。方法:设计leptin基因引物,从人脂肪组织中RT-PCR扩增leptin基因,将获得的基因插入真核表达载体pIRES2-EGFP中,得到重组质粒pIRES2-EGFP-LEP,限制性内切酶双酶切鉴定重组质粒;用脂质体法将重组质粒转染至HPMSCs中并通过RT-PCR及Western blotting法检测目的基因表达,鉴定转染后HPMSCs的多向分化潜能。结果:RT-PCR扩增得到的特异性片段长度约为500 bp,重组质粒经双酶切后显示5.3 kb和500 bp左右的2条片段;转染后的HPMSCs表达leptin基因,保持多向分化潜能。结论:成功构建了瘦素基因真核表达载体,将其转入HPMSCs中并得以表达。Objective：To construct a eukaryotic expression vector of leptin gene,and to transfect it into human placental mesenchy-mal stem cells（HPMSCs）and appraise its expression.Methods：Primers were designed and the leptin gene was obtained by RT-PCR from human adipose tissue.The aimed segments were inserted into the eukaryotic expression vector pIRES2-EGFP,plasmid pIRES2-EGFP-LEP was constructed and identified by restricted enzymatic resection,and then transfected it into HPMSCs by liposome.The ex-pression of leptin in the transfected cells was detected by RT-PCR and Western blotting,the multi-differentiation potential of the cells was indentified.Results：The length of specific fragment was 500 bp,the recombinant plasmid pIRES2-EGFP-LEP presented 5.3 kb and 500 bp bands by restriction enzyme digestion.Leptin gene was expressed in transfected HPMSCs and the transfected HPMSCs maintained multi-directional differentiation potential.Conclusion：The eukaryotic expression vector of leptin can be transfected and ex-pressed in HPMSCs.

关 键 词：真核表达载体 人胎盘间充质干细胞 瘦素 

分 类 号：Q78[生物学—分子生物学]