Cloning and expression of Hemagglutinin gene of influenza H9N2 in Sf9 cell

Influenza virus is one of the most important causes of death due to respiratory diseases in poultry, which causes a lot of damage to the poultry industry worldwide every year. Hemagglutinin protein (HA) is one of the main factors in the pathogenesis of this virus. The aim of this research is cloning and expression of this gene in Sf9 cell using baculovirus.

After isolating of the H9N2 influenza virus genome, the HA gene was amplified by specific primers by RT-PCR and PCR methods and transferred to Bakmid using pFastBac Dual vector. The recombinant HA gene was transferred to the DH10Bac host cell. By transfecting Sf9 cell with recombinant bacmid, the expression of recombinant protein was examined by SDS-PAGE, western blotting and Bradford methods.

The protein obtained from recombinant Bakmid was evaluated using SDS PAGE and its purity was confirmed. The concentration of recombinant protein in the supernatant of Sf9 cells infected with recombinant virus was estimated to be 138µg / ml and confirmed by western blotting.

In the present study, the HA gene was successfully cloned on the PfastBacDual vector. With animal experiments, this protein could be used as a recombinant vaccine candida against influenza H9N2 virus.Key word: Influenza virus, Baculovirus, PfastBacDual vector, Recombinant vaccine