Cloning and Expression of Gumboro VP2 Antigen in Aspergillus niger.

Mohammad Azizi (1, 2) , Bagher Yakhchali (1) , Abdolreza Ghamarian (2) , Somayeh Enayati (2) , Mahvash Khodabandeh (1) , Vahid Khalaj (2)

1 Industrial and Environmental Biotechnology Department
2 Biotechnology Research Center

BACKGROUND: Infectious Bursal Disease Virus (IBDV) causes a highly immunosuppressive disease in chickens and is a pathogen of major economic importance to the poultry industry worldwide. The VP2 protein is the major host-protective immunogen of IBDV and has been considered as a potential subunit vaccine against the disease. VP2 coding sequence was cloned in an inducible fungal vector and the protein was expressed in Aspergillus niger (A. niger). METHODS: Aiming at a high level of expression, a multicopy AMA1-pyrG-based episomal construct driven by a strong inducible promoter, glaA, was prepared and used in transformation of A. niger pyrG-protoplasts. SDS-PAGE and western blot analysis was carried out to confirm the expression of the protein. RESULTS: A number of pyrG (+) positive transformants were isolated and the presence of expression cassette was confirmed. Western blot analysis of one of these recombinant strains using monospecific anti-VP2 antibodies demonstrated the successful expression of the protein. The recombinant protein was also detected by serum obtained from immunized chicken. CONCLUSION: In the present study, we have generated a recombinant A. niger strain expressing VP2 protein intracellulary. This recombinant strain of A. niger may have potential applications in oral vaccination against IBDV in poultry industry.

Keywords: Aspergillus niger; Recombinant proteins; VP2 protein

Domains: Biotechnology; Biotechnology

Mohammad Azizi, Bagher Yakhchali, Abdolreza Ghamarian, Somayeh Enayati, Mahvash Khodabandeh, et al.. Cloning and Expression of Gumboro VP2 Antigen in Aspergillus niger.. Avicenna Journal of Medical Biotechnology, 2013, 5 (1), pp.35-41. ⟨pasteur-00825962⟩