Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (41): 7693-7696.doi: 10.3969/j.issn.1673-8225.2011.41.023

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Construction of eukaryotic expression vecter for human ski gene and observation of its role in promoting cell proliferation

Peng Yan, Li Ping, Liu Ping, Chen Lei, Zhao Xiao-guang, Zhou Yuan-guo   

  1. State Key Laboratory of Trauma, Burns and Combined Injury, Center for Molecular Biology, Research Institute of Surgery and Daping Hospital, Third Military Medical University of Chinese PLA, Chongqing   400042, China
  • Received:2011-04-01 Revised:2011-06-12 Online:2011-10-08 Published:2011-10-08
  • Contact: Zhou Yuan-guo, Researcher, State Key Laboratory of Trauma, Burns and Combined Injury, Center for Molecular Biology, Research Institute of Surgery and Daping Hospital,Third Military Medical University of Chinese PLA, Chongqing 400042, China ygzhou@cta.cq.cn
  • About author:Peng Yan★, Master, State Key Laboratory of Trauma, Burns and Combined Injury, Center for Molecular Biology, Research Institute of Surgery And Daping Hospital,Third Military Medical University of Chinese PLA, Chongqing 400042, China pengyan1006@126. com
  • Supported by:

    An Independent Topic of State Key Laboratory of Trauma, Burns and Combined Injury, No. SKLZZ200805*

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Abstract:

BACKGROUND: c-Ski can promote tissue repair and alleviate scar formation, which is expected to be a new gene therapeutic drugs.
OBJECTIVE: To construct eukaryotic expression plasmid of human ski gene and to investigate its proliferative role in fibroblasts.
METHODS: The ski gene was amplified by polymerase chain reaction (PCR) from human foreskin cultured fibroblast and was sub-cloned into pUC118 with CMV eukaryotic promoter. The recombinant pUC118-Ski plasmids were transfected into rat skin fibroblasts using lipofectamine TM 2000 and then its protein expression and effect of the proliferation were detected.
RESULTS AND CONCLUSION: pUC118-Ski vector was constructed successfully which was confirmed by restriction endonuclease digestion and DNA sequencing. It expressed successfully and significantly improved the effect of the proliferation in transfected fibroblasts. Recombinant eukaryotic expression plasmid pUC118-Ski can improve the proliferation of rat skin fibroblasts.

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