Expression of truncated gE gene of pseudorabies virus (PRV) and primary application in differential diagnosis of PRV vaccination and infection / 生物工程学报
Chinese Journal of Biotechnology
; (12): 526-531, 2004.
Article
en Zh
| WPRIM
| ID: wpr-270093
Biblioteca responsable:
WPRO
ABSTRACT
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%.
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Seudorrabia
/
Porcinos
/
Proteínas Recombinantes
/
Ensayo de Inmunoadsorción Enzimática
/
Proteínas del Envoltorio Viral
/
Vacunación
/
Herpesvirus Suido 1
/
Vacunas contra la Seudorrabia
/
Diagnóstico
/
Diagnóstico Diferencial
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2004
Tipo del documento:
Article