CAJ | 학술논문

背景:骨髓间质干细胞是一类具有多向分化能力的成体干细胞,目前,已有将其作为细胞载体对帕金森病进行治疗的相关报道.目的;构建pDsRed-C1-CDNF真核表达载体,并诱导其在大鼠骨髓间质干细胞中表达.方法:通过RT-PCR的方法从小鼠组织中扩增出CDNF基因片段,并在其两端引入Xho I、BamH I限制性内切酶酶切位点,然后将其克隆至pDsRed-C1真核载体,构建pDsRed-C1-CDNF真核表达载体,并通过Lipofectin2000将其转染至大鼠骨髓间质干细胞中.结果与结论:pDsRed-C1-CDNF真核表达载体经双酶切、单酶切、PCR及测序验证正确,提示已成功构建pDsRed-C1-CDNF真核表达载体并已转染至大鼠骨髓间质干细胞中.
배경:골수간질간세포시일류구유다향분화능력적성체간세포,목전,이유장기작위세포재체대파금삼병진행치료적상관보도.목적;구건pDsRed-C1-CDNF진핵표체재체,병유도기재대서골수간질간세포중표체.방법:통과RT-PCR적방법종소서조직중확증출CDNF기인편단,병재기량단인입Xho I、BamH I한제성내절매매절위점,연후장기극륭지pDsRed-C1진핵재체,구건pDsRed-C1-CDNF진핵표체재체,병통과Lipofectin2000장기전염지대서골수간질간세포중.결과여결론:pDsRed-C1-CDNF진핵표체재체경쌍매절、단매절、PCR급측서험증정학,제시이성공구건pDsRed-C1-CDNF진핵표체재체병이전염지대서골수간질간세포중.
BACKGROUND:Bone marrow mesenchymal stem cells(MSCs)are a kind of adult stem cells with multi-potential differentiation property.At present,it has served as cell carrier for the treatment of Parkinson's disease.OBJECTIVE:To construct pDsRed-C1-CDNF eukaryotic expression vector and induce its expression in rat MSCs.METHODS:CDNF gene was amplified from mouse tissues using RT-PCR,and sequence with Xho I,BamHI restriction enzyme cutting site.The CDNF gene was inserted into the eukaryotic expression vector pDsRed-C1 encoding red fluorescent protein gene.The plasmid pDsRed-C1-CDNF was constructed and transfected into rat bone marrow MSCs.RESULES AND CONCLUSION:The pDsRed-C1-CDNF recombinant plasmid was confirmed by double digestion of Xho I and BamHI restriction enzyme or single digestion of BamHI,and PCR sequence.Results show that the recombinant pDsRed-C1-CDNF eukaryotic expression vector was successfully constructed.