Abstract
Objective To efficiently express NP-M2e fusion protein of influenza A virus A/Jingke/30/95 (H3N2) in E. coli. Methods NP-M2e fusion structure (NM2e) was codon-optimized and pET30a-NM2e recombinant plasmids were constructed. Comparative studies were carried out on the gene expression efficiency, induction temperature and time, purification process and immune reactivity. Results NM2e gene was correctly inserted into pET-30a. SDS-PAGE showed that NM2e could be efficiently expressed in E. coli. The lower temperature (T=25℃) and longer time induction (t=10 h) were necessary for high-level expression of soluble NM2e. The purity of NM2e was up to 90% through two-step purification process with anion-exchange and gel filtration chromatography, and purified NM2e reacted well with serum from NP immunized mice and monoclonal antibody against M2e. Conclusions NM2e with specific immune reactivity can be efficiently expressed and purified from supernatant of E. coli lysate. Key words: Influenza A virus; Viral fusion proteins; Prokaryotic cells; Protein engineering
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