大豆主要过敏原β-伴大豆球蛋白β亚基的抗原表位分析及分段克隆

皮江一; 席俊; 贺梦雪; 李爽

doi:10.13386/j.issn1002-0306.2017.18.018

大豆主要过敏原β-伴大豆球蛋白β亚基的抗原表位分析及分段克隆

河南工业大学粮油食品学院

基金项目:

国家自然科学基金项目(31671778;31301409)；河南省高等学校重点科研项目(16A550001)；

详细信息

作者简介:
皮江一 (1993-) , 男, 硕士研究生, 研究方向:食品免疫学, E-mail:577129671@qq.com。;

席俊 (1977-) , 女, 博士, 副教授, 研究方向:食品免疫学, E-mail:Xijunhnu@163.com。;

中图分类号: Q943.2;TS201.21
计量
- 文章访问数: 217
- HTML全文浏览量: 26
- PDF下载量: 154
出版历程
- 收稿日期: 2017-02-22

Analysis of antigenic epitopes and cloning of soybean major allergen β-conglycinin beta-subunit

College of Food Science and Technology, Henan University of Technology

摘要

摘要: 利用生物信息学软件SWISS-MODEL对大豆主要过敏原β-伴大豆球蛋白β亚基建立出三级结构模型,并根据Disco Tope 2.0服务器预测出该模型的构象表位相关区段,采用分段克隆方式扩增目的基因的3个片段区域,利用PCR技术扩增目的基因全长及3个互相重叠的片段（A、B、C）将其连接至p MD18-T载体,并转化到大肠杆菌感受态JM109。经蓝白斑筛选,以及对重组质粒进行PCR和双酶切验证,测序结果与设计的基因序列一致,成功得到了目的基因片段的阳性克隆子,为β-伴大豆球蛋白β亚基分段表达及抗原区域位置的筛选与鉴定提供参考。
- β-伴大豆球蛋白 /
- β亚基 /
- 生物信息学 /
- 抗原位点 /
- 克隆
Abstract: The model of tertiary of β-conglycinin beta-subunit was constructed by the bioinformation tool SWISS-MODEL, and the conformational epitopes were predicted by Disco Tope 2.0 server.The overlapping fragments of β-conglycinin beta-subunit genes were amplified by PCR and inserted into p MD-18 T vector, then preserved into competent cell of Escherichia coli JM109.The recombinant plasmids were evaluated by blue-white selection, PCR and double enzyme digestion, and DNA sequence analysis showed that the cloned sequence was the very fragment designed. The successful cloning of the genes can provide a reference for the study of the expresstion of β-conglycinin beta-subunit and screening of antigenic epitopes.
- β-conglycinin /
- beta-subunit /
- bioinformation /
- antigenic epitopes /
- clone

HTML全文

参考文献(16)

[1]	ALOK V, SANDEEP K, MUKUL D, et al.A comprehensive review of legume allergy[J].Clinical Reviews in Allergy and Immunology, 2013, 45 (1) :30-46.
[2]	HE L, HAN M, QIAO S, et al.Soybean Antigen Proteins and their Intestinal Sensitization Activities[J].Current Protein and Peptide Science, 2015, 16 (7) :613-621.
[3]	SUN X, SHAN X, YAN Z, et al.Prediction andcharacterization of the linear Ig E epitopes for the major soybeanallergenβ-conglycinin using immunoinformatics tools[J].Food and Chemical Toxicology, 2013, 56 (12) :254-260.
[4]	Maruyama N, Adachi M, Takahashi K, et al.Crystal structures of recombinant and native soybean beta-conglycinin beta homotrimers[J].European Journal of Biochemistry, 2001, 268 (12) :3595-3604.
[5]	MEINLSCHMIDT P, SCHWEIGGERT-WEISZU, BRODE V, et al.Enzyme assisted degradation of potential soy protein allergens with special emphasis on the technofunctionality and the avoidance of a bitter taste formation[J].LWT-Food Science and Technology, 2016, 68:707-716.
[6]	朱婷伟, 陈复生, 布冠好, 等.大豆蛋白过敏原结构与功能的研究进展[J].食品工业科技, 2013, 34 (21) :377-380.
[7]	Coates JB, Medeiros JS, Thanh VH, et al.Characterization of the subunits of beta-conglycinin[J].Archives of Biochemistry&Biophysics, 1985, 243 (1) :184.
[8]	郑树贵, 曹松屹, 孙泽威, 等.β-伴大豆球蛋白β亚基的纯化及免疫活性鉴定[J].大豆科学, 2009, 28 (2) :301-304.
[9]	闫慧丽.大豆主要过敏原Gly m Bd 28K Ig G结合表位的定位研究[D].郑州:河南工业大学, 2016.
[10]	Larralde O G, Martinez R, Camacho F, et al.Identification of hepatitis a virus mimotopes by phage display, antigenicity and immunogenicity[J].Journal of Virological Methods, 2007, 140 (1-2) :49-58.
[11]	Maruyama N, Adachi M, Takahashi K, et al.Crystal structures of recombinant and native soybeanβ-conglycininβhomotrimers[J].Eur J Biochem, 2001, 268 (12) :3595-3604.
[12]	卫会君, 司晓霞, 田怀东.β-伴大豆球蛋白β亚基缺失的大豆遗传资源的发掘[J].科技与创新, 2014 (4) :159.
[13]	杨廷亚, 王芳, 姜平, 等.应用噬菌体展示技术筛选兔出血症病毒抗原模拟表位[J].畜牧兽医学报, 2012, 43 (8) :1281-1286.
[14]	袁水林, 李欣, 陈红兵, 等.噬菌体展示技术定位主要食物过敏原表位的研究进展[J].食品工业科技, 2015, 36 (3) :379-384.
[15]	Wang W, Han J X, Wu Y J, et al.Simultaneous detection of eight food allergens using optical thin-film biosensor chips[J].J Agric Food Chem, 2011, 59 (13) :6889-6894.
[16]	张丹, 鲁会军, 金扩世, 等.猪乙型脑炎病毒E蛋白基因的分段克隆及原核表达[C].中国畜牧兽医学会2010年学术年会——中国兽医临床大会, 2010:303-305.