Expression And Epitopes Mapping Of Structural Proteins Of Hendra And Nipah Virus

Two newly emerged paramyxoviruses, now known as Hendra virus (HeV) and Nipah virus (NiV), were identified in Australia and Malaysia in 1994 and 1999, respectively. The broad species tropisms and the ability to cause fatal disease in both animals and humans have distinguished HeV and NiV from all other known paramyxoviruses, these two viruses have been classified in a separate genus Henipavirus. It has been identified that Flying foxes are the natural host of HeV and NiV. In recent years, both HeV and NiV have continued to emerge and reemerge, The epidemic investigation suggested that the virus could be widely present in the Flying foxes, especially Pteropus lylei in Tailand and Cambodia, which are close to China, moreover, the similar enviorment and Flying foxes species,distribution in the south-east area poese a great threat to pig industry and public health of China, based on the above issues, there is a pressing need for diagnostic technologies on the two virus.In order to prevent their potential outbreak in China, in this study, HeV nucleocapsid protein gene (N), NiV nucleocapsid protein gene (N) and NiV glycoprotein gene (G) were amplified by PCR from the plasmid imported from Australia, and cloned into the pET-28(a) expressing vector, transformed to three diffirent host strains BL21(DE3), BL21(DE3)LysS and JM109(DE3), SDS-PAGE showed that HeV N, NiV N genes were expressed effectively in E.coli JM109(DE3), accounting for 13.8%(NiV N)and 14.6% (HeV N) of total bacteria proteins. Since the NiV G was failed to be expressed, three fragment (GA,GB,GC) of NiV G which contain main antigenic region were fused with GST gene and expressed in E.coli, the final result showed that the GC fragment was expressed. The Western-blot and I-ELISA results indicated that the recombinant HeV N, NiV N and GC protein can be recognized by rabbit anti-NiV serum and rabbit anti-HeV serum, this implied that all...