大肠杆菌O157:H7 Ivy蛋白表位多肽及免疫原性分析

doi:10.16431/j.cnki.1671-7236.2017.01.027

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (1): 194-200.doi: 10.16431/j.cnki.1671-7236.2017.01.027

大肠杆菌O157:H7 Ivy蛋白表位多肽及免疫原性分析

潘艳^1,2, 林敏玲³, 李军^1,2, 杨威^1,2, 陈泽祥^1,2, 谢永平^1,2, 彭昊^1,2, 胡庭俊³

1. 广西兽医研究所, 南宁 530001;
2. 广西兽医生物技术重点实验室, 南宁 530001;
3. 广西大学动物科学技术学院, 南宁 530005

收稿日期:2016-07-06 出版日期:2017-01-20 发布日期:2017-01-19
通讯作者: 李军, 胡庭俊 E-mail:jlee9981@163.com;tingjunhu@gxu.edu.cn
作者简介:潘艳(1978-),女,广西南宁人,副研究员,研究方向:分子生物学,E-mail:panyan413@163.com;林敏玲(1990-),女,广东佛山人,硕士生,研究方向:兽医药理学及毒理学,E-mail:1091979492@qq.com
基金资助:
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）

Antigen Epitope Peptides and Immunogenicity Analysis of E. coli O157: H7 Ivy Protein

PAN Yan^1,2, LIN Min-ling³, LI Jun^1,2, YANG Wei^1,2, CHEN Ze-xiang^1,2, XIE Yong-ping^1,2, PENG Hao^1,2, HU TING-jun³

1. Guangxi Veterinary Research Institute, Nanning 530001, China;
2. Guangxi Key Laboratory of Veterinary Biotechnology, Nanning 530001, China;
3. College of Animal Sciences and Technology, Guangxi University, Nanning 530005, China

Received:2016-07-06 Online:2017-01-20 Published:2017-01-19

摘要/Abstract

摘要：

为评价合成的O157：H7 Ivy146-157多肽作为疫苗抗原的可行性，本试验利用生物信息学方法分析Ivy的二级结构、氨基酸亲水性、蛋白柔性区域、表面可及性和抗原指数，筛选获得1条含有B细胞抗原表位的多肽序列Ivy146-157，进行化学合成。多肽免疫BALB/c小鼠，测定免疫后小鼠的抗体水平及Ivy146-157多肽对小鼠脾淋巴细胞的刺激增殖情况。结果显示，合成的Ivy146-157多肽第3次免疫小鼠后抗体水平达到1：6 400，制备的Ivy146-157多克隆抗体能够识别天然菌株中的目的蛋白；MTT试验结果显示，Ivy146-157免疫后小鼠脾淋巴细胞增殖明显，与对照组差异显著（P<0.05）。结果表明，Ivy146-157多肽能够诱导机体产生体液免疫和细胞免疫反应，为其作为疫苗用抗原的进一步研究奠定了理论基础。

关键词: 肠出血性大肠杆菌O157:H7; 脊椎动物溶菌酶抑制剂; 多肽

Abstract:

To evaluate the feasibility of synthetic polypeptides O157:H7 Ivy146-157 as a vaccine antigen, a bioinformatics method was employed to analyze the secondary structures, hydrophilicity plot, flexibility plot, surface probability plot and antigenic index of the Ivy. A B-cell epitope peptide sequence was identified and synthesized. BALB/c mice were immunized with Ivy146-157, antibody titers and splenic lymphocytes proliferations in mice were tested. The results showed that the antibody titers were 1:6 400 after immunized mice three times. And the native Ivy protein of O157:H7 R14 strain was identified by polyclone antibody Ivy146-157. The MTT assay results indicated the splenic lyphocytes were increased after immunized polypeptides Ivy146-157, and had significant difference with the control group (P<0.05). These results indicated that polypeptides Ivy146-157 could induce a strong humoral and celluar immune respond, and it would provide a theory as a vaccine antigen for further researches.

Key words: EHEC O157:H7; Ivy; polypeptides

中图分类号:

S852.4^＋3

潘艳, 林敏玲, 李军, 杨威, 陈泽祥, 谢永平, 彭昊, 胡庭俊. 大肠杆菌O157:H7 Ivy蛋白表位多肽及免疫原性分析[J]. 《中国畜牧兽医》, 2017, 44(1): 194-200.

PAN Yan, LIN Min-ling, LI Jun, YANG Wei, CHEN Ze-xiang, XIE Yong-ping, PENG Hao, HU TING-jun. Antigen Epitope Peptides and Immunogenicity Analysis of E. coli O157: H7 Ivy Protein[J]. , 2017, 44(1): 194-200.

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大肠杆菌O157:H7 Ivy蛋白表位多肽及免疫原性分析

Antigen Epitope Peptides and Immunogenicity Analysis of E. coli O157: H7 Ivy Protein

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