Abstract: Using Hind Ⅲ enzyme to digest pcDNA3.1/VEGF, the VEGF fragment (575 bp) was obtained. The recombinant transposition vector pFast/VEGF including VEGF gene was constructed and identified with NcoI enzyme reaction. It was transformed into DH10Bac competent cell to recombine baculovirus shuttle vector Bacmid/VEGF. Agrose electrophoresis and PCR amplication were used to determine recombinant vector. The Bacmid/VEGF was transfected into sf-9 cell to screen virus plaque. As a result, agrose electrophoresis observed bacmid DNA band, PCR amplication obtained 575 bp VEGF fragment, virus solution appeared plaque in diluting to 10^-7 concentration(about 50 plaques/well).

Key words: human VEGF, recombinant baculovirus vector, PCR, plaque screening