CAJ | 학술논문

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。
목적：구건수소（leptin）기인진핵표체재체，장기전염지인태반간충질간세포（HPMSCs）중병감정기표체。방법：설계leptin기인인물，종인지방조직중RT-PCR확증leptin기인，장획득적기인삽입진핵표체재체pIRES2-EGFP중，득도중조질립pIRES2-EGFP-LEP，한제성내절매쌍매절감정중조질립；용지질체법장중조질립전염지 HPMSCs 중병통과 RT-PCR급Western blotting법검측목적기인표체，감정전염후HPMSCs 적다향분화잠능。결과：RT-PCR확증득도적특이성편단장도약위500 bp，중조질립경쌍매절후현시5．3 kb화500 bp좌우적2조편단；전염후적HPMSCs표체leptin기인，보지다향분화잠능。결론：성공구건료수소기인진핵표체재체，장기전입HPMSCs중병득이표체。
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.