Evaluation of the immune responses induced by a dna prime protein boost complex against ibdv in chicken

Abstract

Infectious Bursal disease (IBD) is a highly infectious disease of chicken causing severe economic losses to the farming community worldwide. VP2 is the host protective protein with major neutralizing epitopes for inducing both humoral and cell mediated immunity. IBDV field samples were collected and amplified for confirmation of vvIBDV. Further newlinethe positive samples were subjected to RT-PCR for the identification of immunodominant VP2 protein. The resultant 366 bp fragment of the VP2 gene was amplified, cloned and expressed in pRSETA vector with T7 prokaryotic promoter expression. The protein expression was evaluated by induction studies with IPTG and characterized by SDS PAGE and western blotting analysis. The protein was then purified by immobilized nickel affinity column chromatography and the His-tag was removed by Enterokinase Digestion. The resultant 21 kDa recombinant VP2 protein was characterized by SDS-PAGE followed by western blotting. newlineInfectious Bursal disease (IBD) is a highly infectious disease of chicken causing severe economic losses to the farming community worldwide. VP2 is the host protective protein with major neutralizing epitopes for inducing both humoral and cell mediated immunity. IBDV field samples were collected and amplified for confirmation of vvIBDV. Further newlinethe positive samples were subjected to RT-PCR for the identification of immunodominant VP2 protein. The resultant 366 bp fragment of the VP2 gene was amplified, cloned and expressed in pRSETA vector with T7 prokaryotic promoter expression. The protein expression was evaluated by induction studies with IPTG and characterized by SDS PAGE and western blotting analysis. The protein was then purified by immobilized nickel affinity column chromatography and the His-tag was removed by Enterokinase Digestion. The resultant 21 kDa recombinant VP2 protein was characterized by SDS-PAGE followed by western blotting. newline newline