| 猪源大肠杆菌O157:H7鞭毛蛋白抗原表位分析及验证 |
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| 引用本文： | 林敏玲,李军,潘艳,陈泽祥,杨威,胡庭俊. 猪源大肠杆菌O157:H7鞭毛蛋白抗原表位分析及验证[J]. 中国畜牧兽医, 2015, 42(11): 2856-2861. DOI: 10.16431/j.cnki.1671-7236.2015.11.006 |
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| 作者姓名： | 林敏玲 李军 潘艳 陈泽祥 杨威 胡庭俊 |
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| 作者单位： | 1. 广西大学动物科学技术学院, 南宁 530005;2. 广西畜禽疫苗新技术重点实验室, 南宁 530001;3. 广西兽医研究所, 南宁 530001 |
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| 基金项目： | 广西自然科学基金(桂科自2014GXNSFAA118119);广西科研基本业务费专项(桂科专项14-3);广西畜禽疫苗新技术重点实验室专项(13-051-27-A-3) |
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| 摘 要： | 利用生物信息学分析大肠杆菌O157:H7鞭毛蛋白FliC的二级结构及亲水性、抗原指数、柔性区域和表面可能性等指数,预测大肠杆菌O157:H7鞭毛蛋白FliC的潜在B细胞抗原表位,为其致病性研究提供理论基础。利用DNAStar软件Protean程序中Garnier-Robson方法和Chou-Fasman方法分析鞭毛蛋白FliC的α-螺旋、β-折叠、转角区域和卷曲区域,通过Kyte-Doolittle方法、Karplus-Schulz方法、Emini方法和Jameson-Wolf方法分析鞭毛蛋白FliC的亲水性、柔性区域、表面可能性和抗原指数。综合分析得出鞭毛蛋白FliC 63-74、236-247、338-349、460-471、542-553位氨基酸序列为潜在的B细胞优势抗原表位。化学合成法合成优势抗原表位338-349和460-471肽段,免疫BALB/c小鼠3次后,采用ELISA方法验证抗体水平。ELISA结果显示,338-349、460-471肽段具有很强的抗原性,能引起BALB/c小鼠产生高滴度的抗体。
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| 关 键 词： | 大肠杆菌O157:H7 鞭毛蛋白 B细胞抗原表位 免疫 |
| 收稿时间： | 2015-04-13 |
Antigen Epitopes Analysis and Verification of Escherichia coli O157:H7 Flagellin |
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| LIN Min-ling,LI Jun,PAN Yan,CHEN Ze-xiang,YANG Wei,HU Ting-jun. Antigen Epitopes Analysis and Verification of Escherichia coli O157:H7 Flagellin[J]. China Animal Husbandry & Veterinary Medicine, 2015, 42(11): 2856-2861. DOI: 10.16431/j.cnki.1671-7236.2015.11.006 |
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| Authors: | LIN Min-ling LI Jun PAN Yan CHEN Ze-xiang YANG Wei HU Ting-jun |
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| Affiliation: | 1. College of Animal Science and Technology, Guangxi University, Nanning 530005, China;2. Guangxi Key Laboratory of Animal Vaccines and New Technology, Nanning 530001, China;3. Guangxi Veterinary Research Institute, Nanning 530001, China |
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| Abstract: | To understand the pathogenicity of Escherichia coli O157:H7,the secondary structures,hydrophilicity plot,antigenic index,flexibility plot and surface probability plot of the flagellin FliC of Escherichia coli O157:H7 were analyzed by bioinformatics.The α-helix,β-sheet,turn and coil regions of flagellin FliC were analyzed by the methods of Garnier-Robson and Chou-Fasman of DNAStar software,hydrophilicity plot was used the method of Kyte-Doolittle,the flexibility plot was based on the Karplus-Schulz method,surface probability plot and antigenic index were analyzed by the methods of Emini and Jameson-Wolf,respectively.The results showed that 63 to 74,236 to 247,338 to 349,460 to 471 and 542 to 553 amino acid sites of flagellin FliC might be B cell antigen epitopes.338 to 349 and 460 to 471 peptides were synthesized by chemical synthesis,and then subcutaneously immunized BALB/c mice three times.Antibody response was evaluated by ELISA.The result showed that 338 to 349 and 460 to 471 peptides could induce BALB/c mice to produce high titer antibody. |
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| Keywords: | Escherichia coli O157:H7 flagellin B cell antigen epitopes immunity |
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