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Paper Information

Journal:   AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)   JANUARY-MARCH 2013 , Volume 5 , Number 1 (16); Page(s) 35 To 41.
 
Paper: 

CLONING AND EXPRESSION OF GUMBORO VP2 ANTIGEN IN ASPERGILLUS NIGER

 
 
Author(s):  AZIZI MOHAMMAD, YAKHCHALI BAGHER, GHAMARIAN ABDOLREZA, ENAYATI SOMAYEH, KHODABANDEH MAHVASH, KHALAJ VAHID*
 
* MEDICAL BIOTECHNOLOGY DEPARTMENT, BIOTECHNOLOGY RESEARCH CENTER, PASTEUR INSTITUTE OF IRAN, TEHRAN, 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 inAspergillus 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 transform ants 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(s): ASPERGILLUS NIGER, RECOMBINANT PROTEINS, VP2 PROTEIN
 
References: 
 
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