cDNA Cloning, expression and characterization of an allergenic 60s ribosomal protein of almond (prunus dulcis).
Tree nuts, including almond (prunus dulcis) are a source of food allergens often associated with life-threatening allergic reactions in susceptible individuals. Although the proteins in almonds have been biochemically characterized, relatively little has been reported regarding the identity of the allergens involved in almond sensitivity. The present study was undertaken to identify the allergens of the almond by cDNA library approach. cDNA library of almond seeds was constructed in Uni-Zap XR lamda vector and expressed in E. coli XL-1 blue. Plaques were immunoscreened with pooled sera of allergic patients. The cDNA clone reacting significantly with specific IgE antibodies was selected and subcloned and subsequently expressed in E. coli. The amino acids deducted from PCR product of clone showed homology to 60s acidic ribosomal protein of almond. The expressed protein was 11,450 Dalton without leader sequence. Immunoreactivity of the recombinant 60s ribosomal protein (r60sRP) was evaluated with dot blot analysis using pooled and individual sera of allergic patients. The data showed that r60sRP and almond extract (as positive control) possess the ability to bind the IgE antibodies. The results showed that expressed protein is an almond allergen.Whether this r60sRP represents a major allergen of almond needs to be further studied which requires a large number of sera from the almond atopic patients and also need to determine the IgE-reactive frequencies of each individual allergen.
Abolhassani
Mohsen
M
Department of Immunology, Pasteur Institute of Iran, Tehran, Iran. mabolhassani@yahoo.com
Roux
Kenneth H
KH
D000595:Amino Acid Sequence; D052179:Antigens, Plant / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015723:Gene Library; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D008969:Molecular Sequence Data; D021184:Nut Hypersensitivity / Q000276:immunology* / Q000378:metabolism; D027861:Prunus / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012270:Ribosomes / Q000276:immunology; D016415:Sequence Alignment
D052179:Antigens, Plant; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E