[Expression of truncated gE gene of pseudorabies virus (PRV) and primary application in differential diagnosis of PRV vaccination and infection].
With the application of gE gene deleted pseudarabies 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%.
Ni
Jian-Qiang
JQ
National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin 150001, China.
Zhang
Chun-Ling
CL
Tong
Guang-Zhi
GZ
Qiu
Hua-Ji
HJ
Wang
Yun-Feng
YF
Tian
Zhi-Jun
ZJ
D000818:Animals; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay; D011558:Herpesvirus 1, Suid / Q000276:immunology*; D011557:Pseudorabies / Q000175:diagnosis*; D022421:Pseudorabies Vaccines / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology; D013552:Swine; D014611:Vaccination; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology
D022421:Pseudorabies Vaccines; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C089580:glycoprotein E, Suid herpesvirus 1