食品科学 ›› 2009, Vol. 30 ›› Issue (1): 203-206.doi: 10.7506/spkx1002-6630-200901048

• 生物工程 • 上一篇    下一篇

苦荞过敏蛋白全长基因的克隆、表达及免疫学活性研究

张 昕1,崔晓东1,李玉英1,张德梅2,王转花1 ,*   

  1. 1.化学生物学与分子工程教育部重点实验室,山西大学生物技术研究所 2. 太原市红十字血液中心
  • 收稿日期:2008-01-07 修回日期:2008-08-04 出版日期:2009-01-01 发布日期:2010-12-29
  • 通讯作者: 王转花 E-mail:zhwang@sxu.edu.cn
  • 基金资助:

    国家自然科学基金项目(30470178;30671084);山西省自然科学基金项目(2007011077)

Study on Cloning and Expression of A Full-length Gene of Allergenic Protein of Tartary Buckwheat and Its Immunological Activity

ZHANG Xin1,CUI Xiao-dong1,LI Yu-ying1,ZHANG De-mei2,WANG Zhuan-hua1,*   

  1. (1.Key Laboratory of Chemical Biology and Molecular Engineering, Ministry of Education, Institute of Biotechnology,
    Shanxi University, Taiyuan 030006, China ;2.Taiyuan Red-Cross Blood Center, Taiyuan 030001, China)
  • Received:2008-01-07 Revised:2008-08-04 Online:2009-01-01 Published:2010-12-29
  • Contact: WANG Zhuan-hua1 E-mail:zhwang@sxu.edu.cn

摘要:

荞麦属于一种常见的植物性过敏原。本实验在以往研究的基础上,以苦荞种子总RNA 为模板,通过RTPCR和5'-RACE 等方法,扩增、克隆获得苦荞过敏蛋白全长cDNA 序列。分析表明,该序列编码一个由515 个氨基酸残基组成的功能蛋白,并与甜荞过敏性贮藏蛋白的氨基酸序列同源性达到90% 以上。经原核表达及一步亲和纯化后得到纯度较高的重组苦荞过敏蛋白(rTBt)。采用竞争性ELISA 对其免疫活性分析显示,目的蛋白与荞麦过敏病人血清中的IgE 抗体可以特异结合。

关键词: 苦荞, 过敏蛋白, 克隆与表达, 免疫学活性

Abstract:

Buckwheat is a common allergen in plants. In the present study, a full-length cDNA encoding 515 amino acid residues was amplified and cloned from the total RNA of tartary buckwheat by PCR and RACE methods. Its nucleotide sequence has high homology with main allergenic storage protein mRNA in common buckwheat. The recombinant allergenic protein of tartary buckwheat (rTBt) expressed in Escherichia.coli BL21 (DE3) was purified by using affinity chromatography and it can specifically bind to the IgE in patients serum with protein allergy of tartary buckwheat. These results make an important foundation for further study about the mechanism and function of the major allergenic protein in tartary buckwheat.

Key words: tartary buckwheat, allergenic protein, cloning and expression, immunological activity

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