|本期目录/Table of Contents|

[1]田丽花,叶智鸧,温良优,等.副溶血弧菌免疫原性蛋白的鉴定及克隆表达[J].浙江理工大学学报,2013,30(06):881-886.
 TIAN Li hua,YE Zhi cang,WEN Liang you,et al.Identification and Expression of Immunogenic Protein of Vibrio Parahaemolyticus[J].Journal of Zhejiang Sci-Tech University,2013,30(06):881-886.
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副溶血弧菌免疫原性蛋白的鉴定及克隆表达()
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浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]

卷:
第30卷
期数:
2013年06期
页码:
881-886
栏目:
(自科)生物与生命科学
出版日期:
2013-11-10

文章信息/Info

Title:
Identification and Expression of Immunogenic Protein of Vibrio Parahaemolyticus
文章编号:
1673-3851 (2013) 06-0881-06
作者:
田丽花 叶智鸧 温良优 潘建义 赵辅昆
浙江理工大学生命科学学院, 杭州 310018
Author(s):
TIAN Li hua YE Zhi cang WEN Liang you PAN Jian yi ZHAO Fu kun
School of Life Sciences, Zhejiang Sci Tech University, Hangzhou 310018, China
关键词:
副溶血弧菌 外膜蛋白 免疫原性 VP0802
分类号:
Q511
文献标志码:
A
摘要:
运用蛋白质组学及免疫学技术筛选和鉴定副溶血弧菌的外膜蛋白中具有较好免疫原性的外膜蛋白,为防治副溶血弧菌类疾病的亚单位疫苗的研发奠定基础。用灭活的副溶血弧菌免疫ICR小鼠制备抗血清;通过对外膜蛋白进行SDSPAGE和Western Blot分析筛选免疫原性蛋白,经MALDI TOF/TOFMS质谱分析,对免疫原性蛋白进行鉴定。克隆表达免疫原性蛋白VP0802,并对该蛋白的表达条件进行优化,最后纯化蛋白VP0802,并免疫小鼠获得抗血清,通过Western Blot验证蛋白VP0802的免疫原性。结果:免疫小鼠所得到的抗血清效价为 1∶5-000;经免疫印迹分析确定了12种外膜蛋白具有较好免疫原性,其中10种外膜蛋白经质谱分析得到鉴定,分别为LPS assembly protein, VP0802, Maltoporin, Chitoporin, OmpC, OmpA, OmpU, OmpK, VP1243和VP0966。成功构建免疫原性蛋白VP0802的高表达菌株;该菌株在低温低转速的条件下诱导,目的蛋白仍以包涵体形式存在,在37℃,220 r/min ,IPTG终浓度为1 mmol/L的条件下诱导,目的蛋白表达量较高,纯化得到蛋白VP0802并验证了该蛋白的免疫原性。表明在副溶血弧菌的外膜蛋白中存在多种与免疫原性相关的蛋白,且部分蛋白为首次发现具有免疫原性。

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备注/Memo

备注/Memo:
收稿日期: 2013-03-01
基金项目: 国家自然科学基金(31200110);浙江省自然科学基金(Y2010396)
作者简介: 田丽花(1988-),女,山东滨州人,硕士研究生,主要从事生物化学与分子生物学及微生物蛋白质组学研究。
通信作者: 潘建义,Email:panjy@zstu.edu
更新日期/Last Update: 2013-11-12