P-87: Extraction and Detection of Hypervariable Region of VP2 Protein from Infectious Bursal Disease Virus in Escherichia Coli
Objective: Infectious bursal disease (IBD) virus is the causative agent of gumboro disease, a highly contagious disease in young chickens leading to significant economic losses in poultry industry. The causative agent of the disease belongs to the birnaviridae family. IBDV induces immunodeficiency in chicken due to apoptosis in B cells. Currently, IBD vaccines are produced by killed or attenuated viruses. However, the application of these vaccines is problematic, and there are efforts to design new generation vaccines with better properties. VP2 is the major host-protective structural protein of IBDV which is candidate for producing subunit vaccines. The most immunological epitopes of VP2 protein have been characterized in the hypervariable region (named as hvVP2) which is used for the most molecular phylogeny studies and serves to uniquely identify variant, classic, and very virulent IBDV strains. Materials and Methods: At present study, we aim to use Escherichia coli system for production of recombinant form of variable region of VP2 which forms the conformational epitope recognized by virus neutralizing monoclonal antibodies and several mechanical and chemical methods was used for its extraction and detection in SDS-PAGE . Results: Therefore the corresponding part of the gene was cloned in pET26b and transformed in suitable strains of Escherichia coli as final host for expression. According to the results, urea-sonication method is the most appropriate method for extraction of hvVP2 protein from E.coli . Conclusion: Since, hvVP2 has very hy<font><font>drop</font></font>hobic properties; its expression forms inclusion body in the prokaryotic system which is difficult for extraction and detection in SDS-PAGE. So urea_sonication method is highly efficient for detection of hvVP2 into SDS_PAGE.