Infectious bursal disease virus cloning and structural protein (VP2) expression in Escherichia coli

Abstract

Immunosuppressive diseases like infectious bursal disease (IBD) are serious threat to the poultry meat industry. IBD virus consists of two genomic segments in which segment A encodes all structural and non-structural proteins (VP2, VP3, VP4) and segment B encodes RNA dependent RNA polymerase VP1. In IBD the host immune response is due to the VP2 capsid protein of the virus that leads to the immunosuppression of the birds. In the current research the partial VP2 clones were developed using cRNA based reversed genetic system to understand the molecular determinants of infectious bursal disease virus for pathogenic phenotype and virulence. For the expression analysis, the cloning primers were designed for amplification containing NdeI and EcoRI restriction sites for the attachment of restriction enzymes. VP2 of the IBDV was expressed in E. coli cells with the utilization of pET28a expression vector. Expression analysis of bacterial lysate on SDS-PAGE after IPTG induction of 5 hours with 1mM concentration, 30 KDa VP2 protein band was observed. This study suggests the use of recombinant VP2 (rVP2) as a subunit vaccine.

Molecular Characterization and Demographic Study on Infectious Bursal Disease Virus in Faisalabad District

“…Due to the immunosuppressive nature of the disease, it continuously adds to the economic losses in the poultry sector due to compromised cellular and humoral responses after the destruction of B cells resulting in the lower feed conversion ratio. The only way to control IBD is vaccination however, the outbreaks of this disease are also documented in the vaccinated flocks due to the introduction of new virus strains [ 1 ] The possible reason could be the mutation (amino acid changes) that may alter the protection level of the chickens. The genogroups 1, 2 and 3 accommodate the large population of the IBDVs while the classical strains fall in genogroup 1.…”

“…IBD virus is a non-enveloped dsRNA virus with a genome having two segments A and B with a diameter of 55–60 nm. Two serotypes of this virus have been recognized, of which only Serotype-I is known to cause disease in birds [ 1 ]. Serotype-II infects turkeys and several avian species, but no disease has been reported due to it.…”

Advancing IBDV diagnostics: a one-step multiplex real-time qRT-PCR for discriminating between vvIBDV and non-vvIBDV viruses, including the newly emerged IBDV variant

“…The immune suppression caused by IBD leads to diminished responsiveness to commonly used vaccines and increases the impact of other diseases ( 1 ). It is caused by the IBD virus (IBDV), a non-enveloped, double-stranded RNA (dsRNA) virus that belongs to the genus Avibirnavirus under the family Birnaviridae and is composed of two segments (A and B) ( 2 ). Segment A (3.2 kb) is formed from two overlapped open reading frames: the first encodes VP5, while the second encodes the VP2, VP3, and VP4 polyprotein, among which VP2 and VP3 are structural proteins.…”