Expression of M1 and NA Genes of H9N2 Subtype Avian Influenza Virus in Insect Cells
In order to express M1 and NA proteins of H9N2 subtype avian influenza virus(AIV)through insect cells and to identify their immune activity,M1 and NA genes were amplified by PCR,recombinant transfer vectors of pFastBacDual-M1 and pFastBacDual-NA were constructed based on pFastBacDual acting as a recombinant transfer vector;the positive recombinant transfer vectors were respectively transformed into DH10Bac competent cells to obtain recombinant bacmids of rBacmid-M1 and rBacmid-NA;Sf9 insect cells were transfected by the recombinant bacmids to obtain recombinant baculovirus containing M1 and NA genes,that were,rBV-M1 and rBV-NA;the expression of M1 and NA proteins was identified by IFA and Western-blot,and NA protein was used as a coated antigen to identify its reactivity by the indirect ELISA.The results showed that specific green fluorescence was observed by IFA identification,M1 and NA proteins were about 28 and 52 ku,respectively,as tested by Western-blot,and the expressed NA protein could specifically react with H9N2 positive serum.In conclusion,M1 and NA proteins with reactivity could be specifically expressed in insect cells.A foundation was laid for further development of diagnostic techniques and vaccine for H9N2 subtype AIV.
AIVH9N2 subtypeM1 proteinNA proteinBac-to-Bac baculovirus expression system
国家科技期刊平台
NSTL
万方数据
维普
