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
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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184NQEQEF189 7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。Abstract: 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: 27QQN29,39LKPDNRIE46,58NNK60,114PQQKGQSSRP123,134R, 174QMPR177,184NQEQEF189. The size of the target gene matched expectations,the cloning vector was successfully constructed and the phage was successfully assembled. 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.
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Key words:
- glycinin /
- G3A1b polypeptide chain /
- antigenic epitope /
- antigenicity
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