Expression of an epitopic region of AspfI, an allergen/antigen/cytotoxin of Aspergillus fumigatus

The gene for an 18 kD allergen/cytotoxin of Aspergillus fumigatus was cloned in pUC-19 vector and expressed in Escherichia coli JM109. Digestion of this gene with AluI resulted in four fragments of 216bp, 120bp, 39bp and 21bp. These fragments were cloned in the Sma-I site of pUC-19. The recombinants...

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Bibliographic Details
Published in:Immunology letters 1999-12, Vol.70 (3), p.151-155
Main Authors: Sarma, P.V.G.K., Purkayastha, Swagata, Madan, Taruna, Sarma, P.Usha
Format: Article
Language:English
Subjects:
Allergens - genetics
Allergens - immunology
Antigens, Fungal - genetics
Antigens, Fungal - immunology
Antigens, Plant
Asp f1 antigen
Aspergillus fumigatus
Aspergillus fumigatus - immunology
Aspergillusfumigatus
AspfI
Cytotoxins - genetics
Cytotoxins - immunology
Epitope(s)
Epitopes
Escherichia coli
Fungal Proteins - genetics
Fungal Proteins - immunology
Fusion protein
Genes, Fungal
Immunodominant Epitopes
Peptide Fragments - genetics
Peptide Fragments - immunology
plasmid pUC-19
Recombinant Proteins - immunology
Ribonucleases - genetics
Ribonucleases - immunology
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Summary:The gene for an 18 kD allergen/cytotoxin of Aspergillus fumigatus was cloned in pUC-19 vector and expressed in Escherichia coli JM109. Digestion of this gene with AluI resulted in four fragments of 216bp, 120bp, 39bp and 21bp. These fragments were cloned in the Sma-I site of pUC-19. The recombinants thus, generated after transformation in E. coli JM109, were screened using monoclonal antibodies raised against the AspfI. The fusion protein containing 120 bp AluI fragment was recognised by the MoAb indicating presence of epitope(s) in the 120 bp region. The study indicates a viable strategy for identification and expression of an immunologically active domain of a major allergen/antigen of A. fumigatus for the first time.
ISSN:0165-2478
1879-0542
DOI:10.1016/S0165-2478(99)00140-6