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Sequence polymorphism of the group 1 allergen of Bermuda grass pollen
Author(s) -
Chang Zn,
Peng Hj,
WC Lee,
Chen Ts,
Kevin Yiqiang Chua,
Tsai Lc,
Chi Cw,
Han Sh
Publication year - 1999
Publication title -
clinical and experimental allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 154
eISSN - 1365-2222
pISSN - 0954-7894
DOI - 10.1046/j.1365-2222.1999.00523.x
Subject(s) - pichia pastoris , biology , amino acid , complementary dna , gene isoform , recombinant dna , peptide sequence , microbiology and biotechnology , allergen , pollen , escherichia coli , biochemistry , gene , botany , allergy , immunology
Background Cyn d 1, the major allergen of Bermuda grass pollen, consists of a number of isoforms. Objective To examine the extent of sequence variation of Cyn d 1 isoforms at the molecular level. Methods A Bermuda grass pollen λZAP II cDNA expression library was immunoscreened with anti‐Cyn d 1 monoclonal antibodies. The reactive clones were isolated, subcloned into Escherichia coli , and sequenced. Some of them were expressed in the yeast Pichia pastoris to obtain recombinant Cyn d 1 proteins. Results Ten cDNA clones were obtained, all these clones encode the full length of Cyn d 1 protein. Their deduced mature proteins can be grouped into: the long ones with 246 amino acids, and the short ones with 244 amino acids. The last two amino acids (AG) of the long Cyn d 1 are deleted in the short Cyn d 1. The remaining amino acid sequences share more than 98% identity; a total of nine amino acid variations were observed. Two recombinant Cyn d 1 proteins (rCyn d 3–2 and rCyn d 5–4) with three amino acid substitutions showed differential IgE‐binding profiles. Conclusion The present study extended our understanding of the primary structure of isoforms of Cyn d 1.