pDsRed-C1-CDNF真核表达载体在大鼠骨髓间质干细胞中的表达  

Expression of pDsRed-C1-CDNF eukaryotic expression vector in rat bone marrow mesenchymal stem cells

作　　者：张俊[1,2] 牛朝诗[1] 高歌[1] 汤深凤 李静[2] 

机构地区：[1]安徽医科大学附属省立医院神经外科,安徽省合肥市230001 [2]安徽医科大学附属省立医院神经分子生物学与神经再生修复研究室,安徽省合肥市230001

出　　处：《中国组织工程研究与临床康复》2010年第14期2518-2522,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：安徽省人才开发基金项目(2006Z037)~~

摘　　要：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XhoI、BamHI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。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.

关 键 词：保守性多巴胺能神经营养因子 帕金森病 真核表达载体 荧光蛋白 骨髓间质干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]