| 大豆球蛋白A3酸性多肽的原核表达及B细胞表位分析 |
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| 引用本文： | 刘宾,滕达,杨雅麟,王建华. 大豆球蛋白A3酸性多肽的原核表达及B细胞表位分析[J]. 中国食品学报, 2012, 12(3): 13-20 |
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| 作者姓名： | 刘宾 滕达 杨雅麟 王建华 |
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| 作者单位： | 农业部饲料生物技术重点实验室 北京100081 ;中国农业科学院饲料研究所基因室 北京100081 |
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| 基金项目： | 十一五科技支撑计划项目 |
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| 摘 要： | 人工合成大豆球蛋白A3酸性多肽基因cDNA序列,构建原核表达载体pET30a-A3,将重组质粒转化大肠杆菌(E.coli)BL21后用IPTG诱导表达,经His亲和层析纯化,获得纯度约91%融合A3蛋白。Western blotting显示,所获得的A3融合蛋白可与对豆粕过敏的仔猪血清发生免疫反应,表明所得融合蛋白具有免疫活性。利用Lasergene7.0中Protean软件对A3蛋白氨基酸序列进行B细胞抗原表位预测和分析,推测其中97～113、156～160、169～176、186～193、271～284和295～312氨基酸区段是A3酸性多肽的B细胞抗原表位,其中169～176区段的平均抗原指数最高,表位抗原性最强。本结果为进一步研究该蛋白致敏机理及单抗制备奠定了初步基础。
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| 关 键 词： | 大豆球蛋白 A3酸性多肽 原核表达 B细胞表位 |
Prokaryotic Expression and B-cell Eptiopes Prediction of A3 Acidic Polypeptide of Glycinin |
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| Liu Bin , Teng Da , Yang Yalin , Wang Jianhua. Prokaryotic Expression and B-cell Eptiopes Prediction of A3 Acidic Polypeptide of Glycinin[J]. Journal of Chinese Institute of Food Science and Technology, 2012, 12(3): 13-20 |
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| Authors: | Liu Bin Teng Da Yang Yalin Wang Jianhua |
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| Affiliation: | 1,2*(1Key Laboratory of Feed Biotechnology,Ministry of Agriculture,Beijing 100081 2Gene Engineering Laboratory,Feed Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081) |
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| Abstract: | A full-length A3 acidic peptide of glycinin cDNA sequence was synthesized and inserted into the prokaryotic expression vector pET-30a.The A3 fusion protein was expressed in Escherichia coli BL21(DE3) in soluble form with the induction of IPTG.The recombinant protein was purified with His-Bind affinity chromatography with a purity of 91%.Western blotting analysis revealed that the recombinant protein had immunological activity,because it could react with the serum of soybean meal allergic piglet.The potential B-cell epitopes of A3 acidic peptide were predicted using the software Protean of Lasergene 7.0,and located at these regions as 97~113,156~160,169~176,186~193,271~284 and 295~312.The average antigen index(AI) of 169～176 region was highest.The results would be helpful to prepare monoclonal antibody against A3 acidic polypeptide and develop its immunodetection assay. |
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| Keywords: | Glycinin A3 acidic polypeptide prokaryotic expression B-cell epitopes prediction |
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