title: Cloning and Expression of Gumboro VP2 Antigen in Aspergillus niger.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran ; Industrial and Environmental Biotechnology Department, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Azizi
authors.forename: Bagher
authors.initials: B
authors.lastname: Yakhchali
authors.forename: Abdolreza
authors.initials: A
authors.lastname: Ghamarian
authors.forename: Somayeh
authors.initials: S
authors.lastname: Enayati
authors.forename: Mahvash
authors.initials: M
authors.lastname: Khodabandeh
authors.forename: Vahid
authors.initials: V
authors.lastname: Khalaj
keywords: Aspergillus niger; Recombinant proteins; VP2 protein