Author/Authors :
Azizi، Mohammad نويسنده Department of Andrology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran , , Yakhchali، Bagher نويسنده , , Ghamarian، Abdolreza نويسنده Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran , , Enayati، Somayeh نويسنده Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran , , Khodabandeh، Mahvash نويسنده , , Khalaj، Vahid نويسنده Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran ,
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.
