Cloning,Sequencing and Expression of N Gene of Rabies Virus Subclone Strain SRV_9

Genomic RNA was isolated from subcloned rabies virus strain SRV 9 and was used as template for cDNA synthesis of its N gene.The cDNA was amplified by PCR,sequenced and cloned into expression vector pET-28b.The recombinant plasmid was then transoformed into E.coli BL21(DE3).Expression of the protein was induced by IPTG.SDS-PAGE and Western-blotting were performed to analyze the N gene production.Results showed that sequence of the N gene of SRV 9 was completely consistent with its original parental strain SDA.SDS-PAGE results showed that the protein was highly expressed in E.coli .It accounted for 23% of the total proteins.Moleclar weight of the expressed protein was 56 kDa.The expressed protein was specific to antisera against RV by Western-blotting analysis,being presented as a single reaction band.All above demonstrated that the cloned gene was nucleoprotein cDNA of rabies virus and the protein was rabies nucleoprotein specific.It laid a foundation for construction of nucleoprotein genetic vaccine was well for diagnostic reagent development.