Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (18): 3357-3360.doi: 10.3969/j.issn.1673-8225.2011.18.031

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Construction of eukaryotic expression vector carrying rat heat shock protein 72 gene and its expression in COS7 cells

Yan Chun-gen, Huang Dan-wen, Ren Guang-yuan, Zhu Dong-fang   

  1. Department of Internal Medicine, the Affiliated Hospital of Shaoxing University, Shaoxing  312000，Zhejiang Province, China
  • Received:2010-10-22 Revised:2011-03-21 Online:2011-04-30 Published:2011-04-30
  • About author:Yan Chun-gen★, Master, Professor, Department of Internal Medicine, the Affiliated Hospital of Shaoxing University, Shaoxing 312000，Zhejiang Province, China ycg28@163.com
  • Supported by:

    the Natural Science Foundation of Zhejiang Province, No. Y2100961*; Science and Technology Plan of Zhejiang Province, No. 2009C33129*

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Abstract:

BACKGROUND: In vitro cloning and expression of heat shock proteins (HSP), especially HSP72, is important in the study of ischemia-reperfusion injuries.
OBJECTIVE: To construct eukaryotic expression vector carrying HSP72 and study its transient expression in COS7 cells, and to establish the foundation for further research of the immune function of HSP72.
METHODS: HSP72 cDNA was amplified from hepatocytes of BABL/C mice by RT-PCR, and correctly inserted into corresponding sites of eukaryotic expression vector pcDNA3.1(+)after restriction endonuclease digestion, and the recombinant plasmid was transfected into COS7 cells and gene expression was determined by RT-PCR.
RESULTS AND CONCLUSION: The gene fragment inserted into the vector pcDNA3.1(+)was confirmed by nucleotide sequencing, and the recombinant plasmid mRNA was successfully expressed in COS7 cells.

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