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Apple Allergy: The cDNA Sequence of the Major Allergen of Apple, Determined by Performing PCR with a Primer Based on the N ‐Terminal Amino Acid Sequence, is Highly Homologous to the Sequence of the Major Birch Pollen Allergen
Author(s) -
Schöning Britta,
Ziegler Wolfgang H,
Vieths Stefan,
Baltes Werner
Publication year - 1996
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/(sici)1097-0010(199608)71:4<475::aid-jsfa603>3.0.co;2-c
Subject(s) - complementary dna , peptide sequence , allergen , primer (cosmetics) , biology , malus , protein sequencing , amino acid , nucleic acid sequence , microbiology and biotechnology , coding region , genetics , gene , chemistry , botany , allergy , immunology , organic chemistry
Considering the known N‐terminal amino acid sequence of the major apple allergen, a polymerase chain reaction (PCR) primer was selected to amplify cDNA encoding this protein. A single PCR product was obtained, cloned into Escherichia coli and subsequently sequenced. The missing 5′‐end of the apple cDNA sequence was obtained by a 5′‐RACE method. The cDNA sequence showed 72% identity with the coding region of one of the known isoforms of Bet v 1, the major allergen of birch pollen. The deduced amino acid sequence resulted in a 158‐residue protein with a calculated molecular mass of 17·5 kDa and 63% amino acid sequence identity to Bet v 1. In addition, further protein alignments showed a high degree of identity with allergens from other tree pollens and some ‘pathogenesis‐related proteins’ from food plants. According to international regulations the allergen was termed Mal d 1 for this protein, it being the first major allergen discovered and characterised in fruits of apple ( Malus domestica ).