Journal of Capital Medical University ›› 2006, Vol. 27 ›› Issue (5): 623-626.

• 基础研究 • Previous Articles     Next Articles

Construction of Baculovirus Expression System of Human VEGF Gene and Assessment of Biological Activity

Sun Licui1, Wang Yamei1, Yan Yudong2, Zhang Jingyi2, Si Yang2, Qi Yahui2   

  1. 1. Faculty of Biochemistry and Molecular Biology, Capital University of Medical Sciences;2. Medical Experiment and Test Center, Capital University of Medical Sciences
  • Received:2006-01-05 Revised:1900-01-01 Online:2006-10-24 Published:2006-10-24

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Abstract: Objective To construct recombinant baculovirus vector containing VEGF gene and study its biological activity.Methods Using Hind Ⅲ enzyme to digest pcDNA3.1/VEGF,the VEGF fragment(575 bp) was obtained.Then recombinant vector pFast/VEGF including VEGF gene was constructed.It was transformed into DH10BAC competent cell and cultured in LB plate which contained Kan,Ter,Get and X-gal/IPTG.After transfected into sf9 cells the virus plaque was screened.Western blot and MTT methods were used to determine the expression and biological activity of VEGF gene. Results The virus plaque was observed successfully in diluting to 10~(-7) concentration.Western blot showed only one band was identified.Cell growth promotion rate was 19.44% by MTT method.Conclusion The vector pBacmid-Fa-VEGF was successfully constructed and showed significant biological activity to stimulate endothelial cells proliferation.

Key words: human VEGF, recombinant baculovirus vector, Western blot, sf9 cell, MTT

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