Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (37): 6897-6901.doi: 10.3969/j.issn.2095-4344.2012.37.012

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Construction of vascular endothelial growth factor 165 gene eukaryotic vectors for transfection of skeletal muscle cells

Dong Gang1, 2, Zheng Jian-jin2, Li Tao2, Xu Xin1, Lu Shu-lai2, Wu Hong2   

  1. 1Stomatology School, Shandong University, Jinan 250012, Shandong Province, China
    2Qingdao Municipal Hospital, Qingdao 266011, Shandong Province, China
  • Received:2012-01-05 Revised:2012-02-15 Online:2012-09-09 Published:2012-09-09
  • Contact: Zheng Jian-jin, Master, Chief physician, Master’s supervisor, Qingdao Municipal Hospital, Qingdao 266011, Shandong Province, China ZHjj19631016@sina.com
  • About author:Dong Gang☆, Studying for doctorate, Attending physician, Stomatology School, Shandong University, Jinan 250012, Shandong Province, China; Qingdao Municipal Hospital, Qingdao 266011, Shandong Province, China donggangann@sina.com

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Abstract:

BACKGROUND: While researcher became more and more interesting in treating tissue damage by transfection of vascular endothelial growth factor (VEGF), and construction of human VEGF (hVEGF) gene expressive plasmid is of significance.
OBJECTIVE: To clone VEGF gene and to construct the expressive plasmid pcDNA4-HisMax-C/VEGF165, and to observe its expression in rat skeletal muscle cells.
METHODS: hVEGF gene was amplified by reverse transcription-PCR method from human peripheral blood and constructed into the expressive plasmid pcDNA4-HisMax-C/VEGF165. The gene in the expressive plasmid pcDNA4-HisMax-C/VEGF165 was identified by PCR amplification, enzyme digestion and DNA sequencing. pcDNA4-HisMax-C/VEGF165 was transfected into skeletal muscle cells. After 1 week, the VEGF gene cloning was identified.
RESULTS AND CONCLUSION: The cloned DNA was confirmed to be VEGF165 gene. After VEGF165 gene transfection, the cloned DNA from skeletal muscle cells was confirmed to be VEGF165 gene. In this study we successfully cloned VEGF165 gene and constructed its expressive plasmid pcDNA4-HisMax-C/VEGF165, and proved the efficiency of transfection preliminarily.

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