Prokaryotic Expression of F Gene of NDV and Antigenicity of Expressed Product

Newcastle disease virus fusion protein F gene was subcloned into a prokaryotic expression vetor,and then using E.coli BL21(DE3) to express this protein.F gene was amplified from plasmid pMD19T-E2 by PCR,then the gene was subcloned into pET-28a;PCR,digestion and sequencing were used to identify positive plasmid,which named pET-F1;the identified recombined plasmid was expressed by E.coli BL21(DE3) and induced by IPTG.The expressed products were identified by SDS-PAGE and Western blot.The sequence of F1 gene fragment was consistent with predicted size,852 bp.PCR,digestion and sequencing results showed that the recombinant plasmid pET-F1 was correct.Fusion protein F1 was identified by SDS-PAGE and Western blot.Construction of recombinant plasmid pET-F1 containing fusion protein genes,and E2 protein was successful expressed in E.coli BL21(DE3).