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, *
* Corresponding author
1 Industrial and Environmental Biotechnology Department
2 Biotechnology Research Center
Abstract : 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.
keyword : VP2 protein Aspergillus niger Recombinant proteins
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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, Avicenna Research Institute, 2013, 5 (1), pp.35-41. ⟨pasteur-00825962⟩

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