中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (14): 2518-2522.doi: 10.3969/j.issn.1673-8225.2010.14.010　

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

pDsRed-C1-CDNF真核表达载体在大鼠骨髓间质干细胞中的表达

张  俊1,2，牛朝诗1，高  歌1，汤深凤2，李  静2   

  1. 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　
  • 出版日期:2010-04-02 发布日期:2010-04-02
  • 作者简介:张 俊★，男，1986年生，安徽省合肥市人，汉族，安徽医科大学在读硕士，医师，主要从事神经外科的基础与临床研究。 zhangjun.0210@163.com
  • 基金资助:

    安徽省人才开发基金项目(2006Z037)。

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

Zhang Jun 1, 2, Niu Chao-shi1, Gao Ge1, Tang Shen-feng2, Li Jing2   

  1. 1 Department of Neurosrugery, Anhui Provincial Hospital, Anhui Medical University, Hefei   230001, Anhui Province, China; 2 Molecular Neurobiology & Neural Regeneration and Repairing Institute of Anhui Provincial Hospital, Anhui Medical University, Hefei   230001, Anhui Province, China
  • Online:2010-04-02 Published:2010-04-02
  • About author:Zhang Jun★, Studying for master’s degree, Physician, Department of Neurosrugery, Anhui Provincial Hospital, Anhui Medical University, Hefei 230001, Anhui Province, China; Molecular Neurobiology & Neural Regeneration and Repairing Institute of Anhui Provincial Hospital, Anhui Medical University, Hefei 230001, Anhui Province, China zhangjun.0210 @163.com.
  • Supported by:

    the Talent Development Foundation Program of Anhui Province, No. 2006Z037*

PDF

414

被引次数

4

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。

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

Abstract:

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.

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