Construction of prokaryotic expression vector for multiple neutralizing epitopes of human influenza a virus subtype H3N2

Li-Jin-Bo; Zhu-Lei; Qi-Yang

Chinese Journal of Biologicals 21(3): 178-180,189

2008


ISSN/ISBN: 1004-5503
Accession: 030692206

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Abstract
Objective To construct a prokaryotic expression vector for multiple neutralizing epitopes of human influenza A virus subtype H3N2 and express in E. coli Methods The cascaded gene of human influenza A virus subtype H3N2 was synthesized and cloned into vector pET-28a( +). The constructed recombinant plasmid pEr-28a-H3 was identified by restriction analysis and DNA sequencing, then transformed to E. coli BL21(DE3) for expression under induction of IPTG. The expressed product was identified by ELISA and Western blot. Results Both the length and DNA sequence of gene fragment extracted from recombinant plasmid pET-28a-H3 were consistent with those expected. The expressed protein, with a relative molecular mass of about 9 400, showed good reactivity with the corresponding polyclonal antibody. Conclusion The prokaryotic expression vector pEF-28a-H3 for multiple neutralizing epitopes of human influenza A virus subtype H3N2 was successfully constructed and expressed, which laid a foundation of preparation of monoclonal and polyclonal antibodies.