Expression of the Hemagglutinin and Neuramidinase Gene of Influenza A Virus H9N2 in E. coli

Abstract:

To clone, express and characterize the HA and NA Protein of avian influenza A virus H9N2. On the basis of successful clone the full length HA and NA gene and sequence analysis of avian influenza A virus H9N2, we were ligated part of the gene into pET32a (+) and full of the gene into pGEX4T-1. An expression vector pET32a (+)/HA (cut), pET32a (+)/NA (cut), pGEX4T-1/HA, pGEX4T-1/NA were constructed and expressed in E. coli BL21/rosetta induced by IPTG. Recombinant protein was purified through affinity chromatography column. Western Blotting and ELISA were used to determine the antigenic of the recombinant protein. The recombinant capsid gene can be overexpressed in E. coli. SDS-PAGE result showed that the gene could express product as same as we expect. ELISA and Western Blotting result showed that the recombinant protein has good antigenic. The HA and NA protein of avian influenza virus H9N2 has been successful cloned and expressed, which could be useful for developing diagnose reagents or vaccine of H9N2.