71. Infectious bursal disease virus cloning and structural protein (VP2) expression in Escherichia coli
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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.
Keywords: Escherichia coli; IBDV; I-ELISA; RT-PCR; rVP2
http://dx.doi.org/10.19045/bspab.2020.90080