Expression of the hemagglutinin and neuramidinase gene of influenza a virus H9N2 in P. methanolica.

Objective:To clone,express and characterize the HA(hemagglutinin) and NA(neuramidinase,NA) protein of avian influenza virus H9N2.Methods:On the basis of successful clone the full length HA and NA gene and sequence analysis of avian influenza virus H9N2,part of the gene were ligated into pMET A.An expression vector pMETA/HA(52 ～ 1 545bp),pMET A/NA(121 ～ 1 254bp) were constructed and expressed in pMAD16 induced by methanol.Recombinant protein was purified through Ni2 affinity chromatography column.Western blotting and ELISA were used to determine the antigenic of the recombinant protein.Results:The recombinant capsid gene can be overexpressed in P.methanolica.SDS-PAGE showed that the product is a dimer.The purity of the protein is greater than 95%.ELISA and Western blotting showed that the recombinant protein has good antigenic.Conclusion: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.