Prokaryotic expression of main antigen epitope of pseudorabies virus gE gene in Escherichia coli and establishment of indirect ELISA

Objective To distinguish pseudorabies virus (PRV) -infected pigs from those vaccinated with glycoprotein E (gE) -negative vaccine, the method of rapid diagnosis was established.

Method The gE gene fragment of PRV containing the main antigen region was amplified by PCR and cloned into the pro karyotic expression vector pET32a (+) to establish the recombinant plasmid pET-gE184. The recombi nant plasmid was transformed into Escherichia coli BL21 (DE3). After being induced with IPTG, the tar get protein was identified by SDS-PAGE and Western-blot. Wells of ELISA plates were coated with puri fied recombinant protein gE184 to establish the indirect gE184-ELISA.

Result and conclusion Two hundred ninety clinical porcine sera samples have been detected by this ELISA. Compared with commer cial ELISA Kit, the coincidence rate was 93.1%.