Prokaryotic expression and immunogenicity identification of goose tembusu virus E protein domain I.

[Objective] The prokaryotic expression and immunogenicity of goose tembusu virus (GTMUV) E protein domain I were identified in order to lay the foundation for further researches on biological characteristics and immunological function of E protein domain I in GTMUV. [Method] The encoding gene(EI% of GTMUV E protein domain I was artificially synthesised and was inserted into vector pGEX-4t-1 with GST tag. Then the recombinant plasmid was transformed into Escherichia coli. Through induced expression, the fusion protein was obtained and its immunogenicity was identified by using Western blotting. [Result] The obtained gene fragment E1 was 411 bp in length. Through inserting into vector pGEX-4t-1, the recombinant expression plasmid pGEX-4t-1-EI was constructed successfully. When positive recombinant strain was induced with 1 mmol/L IPTG, the expression quantity of fusion protein reached the peak after 5 hours. The fusion protein existed as inclusion body in E. coli, and the molecular weight was about 41.0 kD. The identification result of Western blotting showed that fusion protein could react specifically with GST McAb and E protein positive serum to detect out target bands. [Conclusion] The E protein domain I of GTMUV can be successfully expressed in E. coli, and the obtained fusion protein has good immunogenicity. Therefore it can be used for research & development of GTMUV serology detection kit. [ABSTRACT FROM AUTHOR]