Expression vector construction and prokaryotic expression of the main antigen domains of classical swine fever virus E2 gene

H.H.i; Qiu ChangQing; Zhang YanMing

Chinese Journal of Veterinary Science and Technology 34(1): 3-7

2004


Accession: 004157812

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Abstract
The main antigen domains (A-D) of the E2 gene of CSFV of C-strain, Shimen strain and two prevalent virulent strains (Lintao and Xinjiang strains) were amplified by reverse transcription (RT) and nested polymerase chain reaction (nPCR). The amplified E2 fragments of four strains were all 561 bp in length and 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, size and reading frame were determined by PCR, restriction digestion and the sequence analysis. The prokaryotic expression vectors were constructed and 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 be expressed successfully in E. coli. 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%.