The major allergen (parvalbumin) of codfish is encoded by at least two isotypic genes: cDNA cloning, expression and antibody binding of the recombinant allergens.
The major allergen (parvalbumin) from cod, designated Allergen M Gad c 1, has been intensively studied both from the structural and immunological sides. In the present study, transcripts of two isotypic parvalbumin genes in Atlantic cod were identified and characterized. Subsequently, subfragments were inserted into the expression vector pET-19b, generating plasmids with coding capacity for complete parvalbumin polypeptides fused to an N-terminal his(10) tag. Most of the recombinant products were found in the soluble fraction of the expression host Escherichia coli. The target proteins showed to react with polyclonal antibodies raised against Allergen M and demonstrated binding to specific IgE from 12 sera of patients allergic to cod in ELISA inhibition experiments. Sera with classes 4 and 5 CAP FEIA exhibited also strong binding to recombinant parvalbumins in immunoblots.
Van Do
Thien
T
Department of Clinical Biochemistry, Allergy Research Group, Haukeland University Hospital, University of Bergen, Bergen, Norway.
Hordvik
Ivar
I
Endresen
Curt
C
Elsayed
Said
S
D000485:Allergens; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005396:Fish Products / Q000009:adverse effects; D029941:Fish Proteins / Q000276:immunology; D005399:Fishes / Q000235:genetics* / Q000276:immunology*; D005512:Food Hypersensitivity / Q000276:immunology*; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010320:Parvalbumins / Q000235:genetics* / Q000276:immunology*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D000485:Allergens; D000906:Antibodies; D018076:DNA, Complementary; D029941:Fish Proteins; D010320:Parvalbumins