Abstract: It was to construct a bait vector for vesicular stomatitis virus glycoprotein(VSV-G)and detect its protein expression and self-activation activity in yeast cells, as a basis for further study of screening and VSV-G protein interaction in two hybrid system. Fragment of VSV-G gene were amplified using RT-PCR and directly ligated to pSos vector, insert-contained plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The self-activation and toxicity of the bait plasmid transformed into the yeast cells cdcH25(α)was observed in selective culture medium, and the bait protein was confirmed by Western blot. Results showed that VSV-G gene was found in the reconstructed plasmid pSos-VSV-G by sequencing. Yeast two-hybrid tests showed that yeast cells cdcH25(α)transfected with pSos showed that VSV-G had no self-activation and toxicity, the expression of bait protein was confirmed by Western blot. The yeast two-hybrid system can be applied to screen the interacting proteins of VSV-G.

Key words: Vesicular stomatitis virus, Glycoprotein, Yeast two-hybrid, Bait plasmid