大豆球蛋白G3A1b抗原表位的预测及初步定位  

Prediction and primary mapping of antigenic epitopes of glycinin G3A1b

作　　者：段宇莹 席俊[1] 王一超 付杨 吴枭 孙富宇 陈珍妮 DUAN Yuying;XI Jun;WANG Yichao;FU Yang;WU Xiao;SUN Fuyu;CHEN Zhenni(College of Grain and Oil Food,Henan University of Technology,Zhengzhou 450001,China)

机构地区：[1]河南工业大学粮油食品学院,河南郑州450001

出　　处：《轻工学报》2023年第5期42-50,共9页Journal of Light Industry

基　　金：国家自然科学基金面上项目(32172310);河南工业大学青年骨干教师项目(21420043);河南工业大学创新基金支持计划专项资助项目(2020zkcj19)

摘　　要：采用生物信息学软件对大豆球蛋白中G3亚基A1b酸性多肽链(G3A1b多肽链)的潜在抗原表位进行综合预测,并结合A1b酸性多肽链的氨基酸序列,重叠分段后设计合成3对引物,构建目的基因的克隆载体;利用T7噬菌体展示技术表达目的蛋白,制备热加工特异性吸收抗体后进行ELISA检测以鉴定被破坏的抗原表位。结果表明:G3A1b多肽链中可能存在27QQN29、39LKPDNRIE46、58NNK60、114PQQKGQSSRP123、134R、174QMPR177和184NQEQEF1897个线性表位;目的基因大小与预期相符,成功构建了克隆载体,组装了噬菌体;G3A1b-3片段具有较高的抗原性且在热加工过程中被破坏得最明显。该结果为精确定位大豆球蛋白G3A1b中致敏关键氨基酸提供了理论支持。In this study,b Bioinformatics software was used to predict the potential antigenic epitopes of G3 subunit A1b acidic polypeptide chain in glycinin.Three pairs of primers were designed and synthesized according to the overlapping amino acid sequence of A1b polypeptide chain,and the cloning vector of the target fragment was constructed.The target protein was expressed by phage display technology,and the heat-processed antibody was prepared and the destroyed antigenic epitopes were identified by ELISA.The results showed that G3A1b may contain 7 linear epitopes:27 QQN 29,39 LKPDNRIE 46,58 NNK 60,114 PQQKGQSSRP 123,134 R,174 QMPR 177,184 NQEQEF 189.The size of the target gene was verified by PCR matched expectations,the cloning vector was successfully constructed and the phage was successfully assembled.The results of ELISA showed that G3A1b-3 fragment had high antigenicity and was most obviously destroyed in the process.The results provide theoretical support for the precise positioning of the key amino acids in glycinin G3A1b.

关 键 词：大豆球蛋白 G3A1b多肽链 抗原表位 抗原性 

分 类 号：TS214.2[轻工技术与工程—粮食、油脂及植物蛋白工程]