Cloning and expression of cDNA encoding the complete prepro-form of an isoform of Der f 1, the major group 1 allergen from house dust mite Dermatophagoides farinae.
cDNA clones encoding a major house dust mite allergen, Der f 1, were isolated from a Dermatophagoides farinae cDNA library by plaque immunoscreening using rabbit anti-Der f 1 serum. The sequences cover the complete open reading frame encoding the prepro-form. The sequence is different from previously reported cDNA of Der f 1 in six bases and the encoded amino acid sequence is different in two residues. Pro-forms of Der f 1 and its mutant, in which the N-glycosylation motif was disrupted, expressed in Pichia pastoris were converted to the mature forms by an in vitro activation process and they showed significant IgE-binding. The biologically active rDer f 1 molecules would be useful for diagnostic testing and allergen-specific immunotherapy. In contrast, Der f 1 directly expressed in Escherichia coli without the prosequence had very low IgE binding. The hypoallergenic Der f 1 polypeptide could be useful for safer and more effective immunotherapy.
Yasuhara
T
T
Bioscience Research and Development Laboratory, Asahi Breweries, Ltd., Kitasoma-gun, Ibaraki, Japan.
Takai
T
T
Yuuki
T
T
Okudaira
H
H
Okumura
Y
Y
D000485:Allergens / Q000235:genetics* / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D039741:Antigens, Dermatophagoides; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D015870:Gene Expression; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology; D008925:Mites; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology; D011498:Protein Precursors / Q000235:genetics* / Q000276:immunology; D017386:Sequence Homology, Amino Acid
D000485:Allergens; D039741:Antigens, Dermatophagoides; D018076:DNA, Complementary; D006023:Glycoproteins; D020033:Protein Isoforms; D011498:Protein Precursors; D007073:Immunoglobulin E