Established Of Indirect ELISA Assay With The Prokaryotically Expressed Rabies Virus P Gene Product

In order to established a kind of method to detect the antibody of rabies virus.Amplied rabies virus of LEP-Flury strain by BHK21.The complete length of P gene from rabies virus was amplified by RT-PCR using a pair of specific primers designed according to the relevant sequences from GenBank.The PCR product was cloned into cloning expression vestor pGM-T to obtain the cloning expressed plasmid pGM-T-P. After double-digested by NotI and EcoRI, the product was transferred into prokaryotic expression vetor pET-32a (+) to abstain the prokaryotically expressed plasmid pET-32a-P. The target gene was then expressed in the E.coli BL21(DE3) cell by inducted with IPTG. The highest expression of target protein was analysed by SDS-PAGE, and the good immunoreactivity to rabies virus antibodies was proved by Western-blot analysis. By using purified protein, indirect ELISA for the detection of rabies virus antibodies in canine serum was applied after management of the optional woking condition.