Prokaryotic Expression Of Infectious Bursal Disease Virus Vp2Gene And Immunogenic Detection

In this study,VP2gene of IBDV was cloned from super wild strain HB(hebei) of IBDV of chicken by means of RT-PCR technique,and a fragement of1536bp was amplified from it. And it connects the pMD19-T transformed into competent DH5a E.coli.Comparing with IBDV strain reported in GenBank has been found that B87vaccine strain homology of90%or more after sequencing of IBDV super wild strain (HB).Doing the identification of double enzyme digest after recycling the agarose gel,then VP2has been transfered to the pGEx-4T-1vector and expressed and transferred to the BL21recipient strain that has been induced expression.The results of SDS-PAGE and Western blot indicated that the expressed VP2protein of molecular weight was about66kDa.The recombinant protein was purified by GST agarose gel affinity chromatography and was made into subunit vaccine by white oil adjuvant that has been emulsionized.Compared with the inactive vaccine of IBDV from different manufacture in order to test the immunogenicity of recombinant expression pGEx-4T-1-VP2protein.Chicken were challenged with a virulent IBDV stain (HB) by oral and eye drops,the results show that protection rate of commercialize vaccine group and recombinant expression pGEx-4T-1-VP2protein group were100%,50%,respectively...