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Cloning and sequencing of the Bet v 1-homologous allergen Fra a 1 in strawberry (Fragaria ananassa) shows the presence of an intron and little variability in amino acid sequence

Musidlowska Persson, Anna LU ; Alm, Rikard LU and Emanuelsson, Cecilia LU (2007) In Molecular Immunology 44(6). p.1245-1252
Abstract
The Fra a I allergen in strawberry (Fragaria ananassa) is homologous to the major birch pollen allergen Bet v 1, which has numerous isoforms differing in terms of amino acid sequence and immunological impact. To map the extent of sequence differences in the Fra a I allergen, PCR cloning and sequencing was applied. Several genomic sequences of Fra a 1, with a length of either 584, 591 or 594 nucleotides, were obtained from three different strawberry varieties. All contained one intron, with the length of either 10 1 or 110 nucleotides. By sequencing 30 different clones, eight different DNA sequences were obtained, giving in total five potential Fra a I protein isoforms, with high sequence similarity (> 97% sequence identity) and only... (More)
The Fra a I allergen in strawberry (Fragaria ananassa) is homologous to the major birch pollen allergen Bet v 1, which has numerous isoforms differing in terms of amino acid sequence and immunological impact. To map the extent of sequence differences in the Fra a I allergen, PCR cloning and sequencing was applied. Several genomic sequences of Fra a 1, with a length of either 584, 591 or 594 nucleotides, were obtained from three different strawberry varieties. All contained one intron, with the length of either 10 1 or 110 nucleotides. By sequencing 30 different clones, eight different DNA sequences were obtained, giving in total five potential Fra a I protein isoforms, with high sequence similarity (> 97% sequence identity) and only seven positions of amino acid variability, which were largely confirmed by mass spectrometry of expressed proteins. We conclude that the sequence variability in the strawberry allergen Fra a I is small, within and between strawberry varieties, and that multiple spots, previously detected in 2DE, are presumably due to differences in post-translational modification rather than differences in amino acid sequence. The most abundant Fra a I isoform sequence, recombinantly expressed in Escherichia coli after removal of the intron, was recognized by IgE from strawberry allergic patients. It cross-reacted with antibodies to Bet v I and the homologous apple allergen Mal d 1 (61 and 78% sequence identity, respectively), and will be used in further analyses of variation in Fra a 1-expression. (c) 2006 Elsevier Ltd. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
isoform, mass spectrometry, cross-reactivity, birch pollen, allergen, allergy
in
Molecular Immunology
volume
44
issue
6
pages
1245 - 1252
publisher
Pergamon
external identifiers
  • wos:000242949700021
  • scopus:33748777681
ISSN
1872-9142
DOI
10.1016/j.molimm.2006.06.004
language
English
LU publication?
yes
id
64361917-cc68-4ab9-8108-bdada002f874 (old id 681477)
date added to LUP
2007-12-13 16:06:37
date last changed
2017-09-24 04:30:12
@article{64361917-cc68-4ab9-8108-bdada002f874,
  abstract     = {The Fra a I allergen in strawberry (Fragaria ananassa) is homologous to the major birch pollen allergen Bet v 1, which has numerous isoforms differing in terms of amino acid sequence and immunological impact. To map the extent of sequence differences in the Fra a I allergen, PCR cloning and sequencing was applied. Several genomic sequences of Fra a 1, with a length of either 584, 591 or 594 nucleotides, were obtained from three different strawberry varieties. All contained one intron, with the length of either 10 1 or 110 nucleotides. By sequencing 30 different clones, eight different DNA sequences were obtained, giving in total five potential Fra a I protein isoforms, with high sequence similarity (> 97% sequence identity) and only seven positions of amino acid variability, which were largely confirmed by mass spectrometry of expressed proteins. We conclude that the sequence variability in the strawberry allergen Fra a I is small, within and between strawberry varieties, and that multiple spots, previously detected in 2DE, are presumably due to differences in post-translational modification rather than differences in amino acid sequence. The most abundant Fra a I isoform sequence, recombinantly expressed in Escherichia coli after removal of the intron, was recognized by IgE from strawberry allergic patients. It cross-reacted with antibodies to Bet v I and the homologous apple allergen Mal d 1 (61 and 78% sequence identity, respectively), and will be used in further analyses of variation in Fra a 1-expression. (c) 2006 Elsevier Ltd. All rights reserved.},
  author       = {Musidlowska Persson, Anna and Alm, Rikard and Emanuelsson, Cecilia},
  issn         = {1872-9142},
  keyword      = {isoform,mass spectrometry,cross-reactivity,birch pollen,allergen,allergy},
  language     = {eng},
  number       = {6},
  pages        = {1245--1252},
  publisher    = {Pergamon},
  series       = {Molecular Immunology},
  title        = {Cloning and sequencing of the Bet v 1-homologous allergen Fra a 1 in strawberry (Fragaria ananassa) shows the presence of an intron and little variability in amino acid sequence},
  url          = {http://dx.doi.org/10.1016/j.molimm.2006.06.004},
  volume       = {44},
  year         = {2007},
}