Yeast expression vector construction of herpes simplex virus type I glycoprotein D

Objective:To construct the cloning and yeast expressing vectors containing HSV-I gD gene and lay foundatio n for expressing the recombinant gD.Methods:The whole sequence of gD was ampl ified from HSV-I DNA genome by PCR,and after being recycled the products were cloned in-to the pMD-18T.Through transformation,conformation and sequence analysis,the cloning vector pMD18T-gD was constructed.The cloning vector a nd the expressing plasmid were digested by restriction enzymes,connected and tr ansformed into E.coli JM109.The products were confirmed using PCR,en-zyme digestion and sequence analysis,and pGAPZaA-gD was constructed.Results:Th e PCR and en-zyme digestion results of both recombinant plasmid pMD18T-gD and pGAPZaA-gD showed the correct fragments,and the sequence analysis confirm ed that the nucleotide sequence was highly conserved,which homology was99.6 6%.Conclusion:The recombinant yeast expression vector of HSV-I gD was cons tructed successfully.