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编码花生过敏相关抗原的克隆的分离、鉴定及特性分析。

Isolation, identification, and characterization of clones encoding antigens responsible for peanut hypersensitivity.

作者信息

Burks A W, Cockrell G, Stanley J S, Helm R M, Bannon G A

机构信息

Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, USA.

出版信息

Int Arch Allergy Immunol. 1995 May-Jun;107(1-3):248-50. doi: 10.1159/000236993.

Abstract

Peanut allergy is a significant health problem because of the frequency, the potential severity, and the chronicity of the allergic sensitivity. Serum IgE from patients with documented peanut hypersensitivity reactions and a peanut cDNA expression library were used to identify clones that encode peanut allergens. One of the major peanut allergens, Ara h I, was selected from these clones using Ara h I-specific oligonucleotides and polymerase chain reaction technology. The Ara h I clone identified a 2.3-kb mRNA species on a Northern blot containing peanut poly A+RNA. DNA sequence analysis of the cloned inserts revealed that the Ara h I allergen has significant homology with the vicilin seed storage protein family found in most higher plants. The isolation of the Ara h I clones allowed the synthesis of this protein in Escherichia coli cells and subsequent recognition of this recombinant protein in immunoblot analysis using serum IgE from patients with peanut hypersensitivity. With the production of the recombinant peanut protein it will now be possible to address the pathophysiologic and immunologic mechanisms regarding peanut hypersensitivity reactions specifically and food hypersensitivity in general.

摘要

花生过敏是一个严重的健康问题,这是由于过敏反应的频繁发生、潜在的严重性以及过敏敏感性的慢性特征。利用有记录的花生过敏反应患者的血清IgE和花生cDNA表达文库来鉴定编码花生过敏原的克隆。使用Ara h I特异性寡核苷酸和聚合酶链反应技术,从这些克隆中筛选出一种主要的花生过敏原Ara h I。该Ara h I克隆在含有花生多聚腺苷酸加尾RNA的Northern印迹上鉴定出一种2.3kb的mRNA。对克隆插入片段的DNA序列分析表明,Ara h I过敏原与大多数高等植物中发现的豌豆球蛋白种子储存蛋白家族具有显著同源性。Ara h I克隆的分离使得该蛋白能够在大肠杆菌细胞中合成,并在使用花生过敏患者血清IgE的免疫印迹分析中识别这种重组蛋白。随着重组花生蛋白的产生,现在有可能专门针对花生过敏反应以及一般的食物过敏反应,研究其病理生理和免疫机制。

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