Prokaryotic and eucaryotic expression of H9N2 avian influenza virus NS1 gene.

【Objective】The study was to construct the prokaryotic expression vector pET-32a-NS1 and eukaryotic expression vector pEGFP-C1-NS1 for the expression of the NS1gene of H9N2 avian influenza virus in E.coli and 293cells.【Method】NS1 gene was amplified by RT-PCR.The NS1 gene was cloned directionaly into the pET-32a(+).The recombinant prokaryotic expression plasmid was constructed and transformed into E.coli BL21(DE3),and induced by IPTG;The NS1 gene was cloned into pEGFP-C1.The recombinant eukaryotic expression plasmid was constructed and transfected into 293 cells by Lipofectami-neTM2000,and the transfected cells were collected 48hours after transfection.Meanwhile,the expressed products were analyzed with Western-blotting.【Result】Results showed that the prokaryotic expression vector and eukaryotic expression vector of NS1gene were successfully constructed,the special band of NS1 protein expressed in E.coli and 293 cells were identified by Western-blotting.【Conclusion】NS1gene wassuccessfully expressed in E.coli and 293cells,Western-blotting test showed that the NS1proteins had very good antigenicity.