[Prokaryotic expression, purification and identification of the recombinant gp90 protein of Reticuloendotheliosis virus].

OBJECTIVE: To obtain the recombinant gp90 protein of Reticuloendotheliosis virus (REV) and the anti-gp90 serum with high titer. METHODS: Using the plasmid pMD18T-env as template, we amplified the gp90 gene and then cloned it into pET-28a(+). The recombinant plasmid pET28a-gp90 was transformed into Escherichia coli BL21 (DE3), which was induced with isopropylthio-beta-D-galactoside(IPTG). After identification by SDS-PAGE and Western blotting, the purified gp90 protein was injected into Balb/c mice to prepare anti-gp90 serum. The specificity and titer of the antiserum were evaluated by IFA and the enzyme-linked immunosorbant assay (ELISA). RESULTS: SDS-PAGE and Western blotting showed that the gp90 protein was expressed successfully in the form of inclusion body in the recombinant E coli. ELISA showed the mouse anti-gp90 serum had a titer of 1:12800. Successful expression of recombinant gp90 protein and preparation of its antiserum laid the foundation for the development of diagnostic reagent of REV.