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无色草莓突变体中草莓Bet v 1同源过敏原的下调与类黄酮生物合成途径协同作用。

Down-regulation of the strawberry Bet v 1-homologous allergen in concert with the flavonoid biosynthesis pathway in colorless strawberry mutant.

作者信息

Hjernø Karin, Alm Rikard, Canbäck Björn, Matthiesen Rune, Trajkovski Karin, Björk Lars, Roepstorff Peter, Emanuelsson Cecilia

机构信息

Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.

出版信息

Proteomics. 2006 Mar;6(5):1574-87. doi: 10.1002/pmic.200500469.

Abstract

Proteomic screening of strawberry (Fragaria ananassa) yielded a 58% success rate in protein identification in spite of the fact that no genomic sequence is available for this species. This was achieved by a combination of MALDI-MS/MS de novo sequencing of double-derivatized peptides and indel-tolerant searching against local protein databases built on both EST and full-length nucleotide sequences. The amino acid sequence of a strawberry allergen, homologous to the well-known major birch pollen allergen Bet v 1, was partially determined. This strawberry allergen, named Fra a 1 according to the nomenclature for allergen proteins, showed sequence identity of 54 and 77%, respectively, with corresponding allergens from birch and apple. Differential expression, as evaluated by 2-D DIGE, occurred in 10% of protein spots when red strawberries were compared to a colorless (white) strawberry mutant. White strawberries, known to be tolerated by individuals affected by allergy, were found to be virtually free from the strawberry allergen. Also several enzymes in the pathway for biosynthesis of flavonoids, to which the red color pelargonidin belongs, were down-regulated. This approach to assess differential protein expression without access to genomic sequence information can also be applied to other crop plants and phenotypic traits.

摘要

尽管草莓(凤梨草莓)没有基因组序列,但蛋白质组学筛选在蛋白质鉴定方面仍取得了58%的成功率。这是通过双衍生肽的基质辅助激光解吸电离串联质谱(MALDI-MS/MS)从头测序与针对基于EST和全长核苷酸序列构建的本地蛋白质数据库进行耐插入缺失搜索相结合实现的。部分确定了一种草莓过敏原的氨基酸序列,该序列与著名的主要桦树花粉过敏原Bet v 1同源。根据过敏原蛋白的命名法,这种草莓过敏原被命名为Fra a 1,与来自桦树和苹果的相应过敏原的序列同一性分别为54%和77%。通过二维差异凝胶电泳(2-D DIGE)评估,当将红色草莓与无色(白色)草莓突变体进行比较时,10%的蛋白质斑点出现了差异表达。已知对过敏个体耐受的白色草莓几乎不含草莓过敏原。此外,红色天竺葵色素所属的类黄酮生物合成途径中的几种酶也被下调。这种在没有基因组序列信息的情况下评估差异蛋白质表达的方法也可应用于其他作物和表型性状。

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