论文部分内容阅读
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-directional differentiation ability. At present, they have been reported as cell carriers for the treatment of Parkinson’s disease. Objective: To construct pDsRed-C1-CDNF eukaryotic expression vector and induce its expression in rat bone marrow mesenchymal stem cells. Methods: The CDNF gene fragment was amplified from mouse tissues by RT-PCR. XhoI and BamHI restriction enzyme sites were introduced into both ends of the CDNF gene, and then cloned into pDsRed-C1 eukaryotic vector. The pDsRed-C1-CDNF eukaryotic expression vector was constructed and transfected into rat bone marrow mesenchymal stem cells by Lipofectin2000. RESULTS AND CONCLUSION: The eukaryotic expression vector pDsRed-C1-CDNF was verified by double enzyme digestion, single enzyme digestion, PCR and sequencing. The eukaryotic expression vector pDsRed-C1-CDNF was successfully constructed and transfected into rat bone marrow In stem cells.