《植物生理学报》 2009, 45(10): 958-962

花生过敏原iso-Ara h 3 的克隆、表达和免疫学鉴定

钟海峰^1,2, 马三梅¹, 王永飞¹, 邹泽红², 陶爱林^2,*

1 暨南大学生物工程学系, 广州510632; 2 广州医学院第二附属医院过敏反应实验室, 广州510260

通信作者：陶爱林；E-mail: AerobiologiaTao@163.com；Tel: 020-34153520

摘　要：

采用 Touchdown PCR 技术从花生 cDNA 中克隆到花生的过敏原 iso-Ara h 3 基因, 并进行重组蛋白表达, 再用 Western blot 技术鉴定重组蛋白过敏原性。结果显示, 构建的 pET44a-iso-Ara h 3 重组菌能表达 iso-Ara h 3 蛋白。用 8 例花生过敏 的阳性血清鉴定表明, 重组的 iso-Ara h 3 蛋白血清 IgE 识别率为 12.5%, 是一种低过敏原性的过敏原蛋白。

关键词：花生; 过敏原; 基因克隆; iso-Ara h 3基因; 过敏原性

收稿：2009-05-31   修定：2009-09-11

资助：国家自然科学基金(30771240)和广东省科技计划(2005B33801002)。

Cloning, Expression and Immunological Characterization of Peanut (Arachis hypogaea L.) Allergen Gene iso-Ara h 3

ZHONG Hai-Feng^{1, 2}, MA San-Mei¹, WANG Yong-Fei¹, ZOU Ze-Hong², TAO Ai-Lin^2,*

¹Department of Biotechnology, Jinan University, Guangzhou 510632, China; ²The Allergy Laboratory, The Second Affiliated Hospital, Guangzhou Medical College, Guangzhou 510260, China

Corresponding author: TAO Ai-Lin; E-mail: AerobiologiaTao@163.com; Tel: 020-34153520

Abstract:

The peanut (Arachis hypogaea) allergen gene iso-Ara h 3 was cloned from the peanut cDNA by using Touchdown PCR. The recombinant iso-Ara h 3 protein was expressed in prokaryotic expression vector. And then the antigenicity of recombinant iso-Ara h 3 proteins was identified by Western blot. The result showed that the recombinant plasmid pET44a-iso-Ara h 3 was efficient and the recombinant iso-Ara h 3 was recongnized by 12.5% of peanut allergic patients. It was concluded that the iso-Ara h 3 allergen exhibited lower allergenicity than other peanut allergens.

Key words: peanut (Arachis hypogaea); allergen; gene cloning; iso-Ara h 3 gene; allergenicity