Expression of Truncated gE Gene of Pseudorabies Virus(PRV) and Primary Application in Differential Diagnosis of PRV Vaccination and Infection
伪狂犬病病毒gE基因主要抗原表位区的原核表达及其在疫苗接种和自然感染鉴别诊断中的应用

With the application of gE gene deleted pseudorabies virus(PRV) vaccine worldwide, a corresponding differential diagnosis based on gE glycoprotein is needed in the project of PRV eradication. In this study, PRV gE gene without signal and transmembrane region was amplified by PCR and cloned into pGEX 6P 1, generated pGEX gE. After transformation of BL21 with pGEX gE, an expressed fusion protein(about 63kD) was identified by SDS PAGE. The recombinant proteins are produced as inclusion bodies. By changing the inductive conditions, the formation of inclusion bodies was inhibited and tended to increase the percentage of soluble recombinant protein. The antigenic reactivity of the recombinant protein was confirmed by Western blotting with polyclonal antibodies against PRV. Using purified recombinant tgE as antigen, an ELISA was developed to detect sera of PRV infected pigs and sera of pigs immunized with gE deleted PRV vaccine. The total of 400 serum samples collected from field were comparatively tested with the tgE ELISA and a commercial competitive ELISA based on monoclonal antibody against gE, the results indicated that the coincidental rate between the two tests is about 94%.