Title: Expression vector construction and prokaryotic expression of the main antigen domains of classical swine fever virus E2 gene
Abstract: The main antigen domains(A-D) of E2 gene of CSFV of C-strain, Shimen strain and two prevalent virulent strains(Lintao strain aod Xinjiang strain) were amplified by reverse transcription(RT) and the nested polymerase chain reaction (nPCR). The amplifed E2 fragments of four strains were all 561bp in length. And they were ligated with plasmid pGEM-T Easy. The recombinant plasmids and expression vector pGEX-4T-1 were digested by the same restriction endonucleases. These genes were ligated and transformed into Escherichia coli. The insert position , the size and the reading frame all were right by PCR, restriction digestion and the sequence analysis. The result showed that the prokaryotic expression vectors were constructed. Then the recombinants were transformed into BL21(DE3) for E2 expression with IPTG inducing. The expressed proteins were measured by SDS-PAGE and Western blotting. The results showed that the E2 genes can express successfully inE.coli. The Western blotting results indicated that the expressed antigen proteins could be recognized by the positive serum of CSFV. The (rate) of the expressed protein in the induced bacteria protein was about 20.1%.
Publication Year: 2004
Publication Date: 2004-01-01
Language: en
Type: article
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