Genomic organization of peanut allergen gene, Ara h 3

被引:16
|
作者
Viquez, Olga M. [1 ]
Konan, Koffi N. [1 ]
Dodo, Hortense W. [1 ]
机构
[1] Alabama A&M Univ, Dept Food & Anim Sci, Food Biotechnol Lab, Normal, AL 35762 USA
关键词
Ara h 3; cis-element; introns; peanut; promoter;
D O I
10.1016/j.molimm.2004.06.033
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Type 1 hypersensitivity to peanut proteins is a well-recognized health problem. Several peanut seed storage proteins have been identified as allergens. Ara h 3, a glycinin protein, is one of the important peanut allergens. Although amino acid and cDNA sequences are available for Ara h 3, there is not information at the genomic level. The objectives of this study were to isolate, sequence, and characterize the genomic clone of peanut allergen, Ara h 3. A peanut genomic library was screened, using two [P-32] end-labeled oligonucleotide probes designed based on cDNA sequences of Ara h 3 and Ara h 4. Four positives lambda FIX II clones were obtained after four rounds of screenings. Digestion with Sac I resulted in two fragments of 1.5 and 10 kb hybridizing to the probes. Both fragments were subcloned into p-Bluescript vector and sequenced. The Ara h 3 gene spans 3.5 kb and consists of four exons, three introns, 5' and 3' flanking regions. The open reading frame is 2008 bp long and can encode a polypeptide of 538 amino acids residues. Sequences analogous to a TATA-box (TATAAAT), CAAT-box (AGGA), G-box (TCCTACGTGTCC) and several cis-elements were found in the promoter region. In the 3' downstream region, three polyadenylation signals (AATAAA) were identified. (C) 2004 Elsevier Ltd. All rights reserved.
引用
收藏
页码:1235 / 1240
页数:6
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