Abstract:ObjectiveTo express gB gene of pseudorabies virus fractionally for developing an ELISA on PRV vaccine antibodies. MethodWe analysed the gB epitope, and designed four pairs of specific primers, then four fragments of gB gene were amplified from PRV Xinjiang strain, which were named for gB-1、gB-3、gB-4 and gB-5 gene. The four gene fragments were cloned into PGEM-T-easy and sequenced, then the four genes were inserted into pGEX-6P-1 vector. The expressed proteins were analyzed by Western blotting. Three indirect ELISA methods were set up using purified gB-3、gB-4 and gB-5 proteins as antigen, and 22 clinical pig sera samples were detected. ResultsThe four recombinant expression plasmids were successfully expressed in Coli, and the proteins molecular weight were 75.05、53.86、49.96 and 49.72×103Da, and gB-3、gB-4 and gB-5 have higher level expression, and the expression product mainly exists in inclusion body. Western blotting identification displayed that all the four fusion proteins were distinguished by pseudorabies virus positive serum. The coincidence rate between gB-3-ELISA, gB-4-ELISA, gB-5-ELISA and gB-ELISA of IDEXX company were respectively 40.9%, 81.8% and 81.8%. ConclusionThe study successfully expressed gB-1、gB-3、gB-4 and gB-5 of PRV gB, gB-3、gB-4 and gB-5 had higher expression. The gB-4-ELISA and gB-5-ELISA had high level coincidence with gB-ELISA of IDEXX company.