Prokaryotic expression and subcloning of gE gene in porcine pseudorabies virus

To develop serological assay for PRV gE-ELISA.Using apair of primers designed according to the revelant nucleotide sequence from GenBank,the specific genes encoding the main antigenic domains of PRV-gE were amplified.The PCR products of 750bp were cloned into the pMDTM19-T vectors.The plasmid pMD-gE'was constructed success-fully.The main antigen of PRV-gEwas cut with BamHⅠand HindⅢ,and linked with prokary-otic expression vector pET32awhich had been cut by the two enzymes before.The plasmid construced was named pET32a-gE'.SDS-PAGE detected that the protein bands was about 45ku,after the recombinant plasmid was induced by IPTG.Western blot analysis showed that,the expression product has goodreactogenicity.