Abstract:

It was to express the Fructose Valine Oxidase Gene with baculovirus expression system. FVO gene was artificially synthesized, and ligated to the vector pFastBac1 to obtain the recombinant vector, which was then transformed into E.coli DH10Bac competent cells to transposase with bacmid. After screening, the recombinant baculovirus Bacmid-pFastBac1-FVO was got, which appeared a single band after PCR identification. sf9 cells was transfected with the recombinant vector, and harvested the virus and expressed the protein. Results showed that the baculovirus expression vector was successfully constructed and expressed in the sf9 cells which appeared a 40 kD band with Western blot.

Key words: Fructose, Valine oxidase, Baculovirus expressive system, Homologous recombination, Sf9 cell, Eukaryotic expression