创伤弧菌外膜蛋白OmpU的基因克隆与表达

李素一; 吴唯维; 李盼; 柯翎; 许斌福; 林晨韬; 林天龙; 陈叙

doi:10.19303/j.issn.1008-0384.2013.06.001

创伤弧菌外膜蛋白OmpU的基因克隆与表达

doi: 10.19303/j.issn.1008-0384.2013.06.001

1. 福建省农业科学院生物技术研究所,福建福州,350003;
2. 福建师范大学,福建福州350007;
3. 福建农林大学,福建福州 350002

基金项目:

福建省自然科学基金项目(2011J05055)

福建省农业科学院青年科技人才创新基金项目(2011QC-20)

国家自然科学基金项目(31100658)

福建省农业科学院引进海外人才启动基金项目(HWRC2011-03)

详细信息

作者简介:
李素一(1985-),女,硕士,研究实习员,主要从事鱼类免疫学研究;陈叙(1978-),女,博士,副研究员,主要从事水产保护学研究(E-mail:chenxu97@hotmail.com)

中图分类号: S941
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出版历程
- 收稿日期: 2013-05-01
- 刊出日期: 2013-06-18

Gene Cloning and Expression of Outer Membrane Protein OmpU of Vibrio vulnificus

1. Biotechnology Institute,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350003,China;
2. Fujian Normal University,Fuzhou,Fujian 350007,China;
3. Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China

摘要

摘要: 创伤弧菌是导致鳗鲡产生体表溃疡死亡的重要致病菌。本研究克隆了创伤弧菌FJ03-X2株外膜蛋白基因(ompU),构建含该基因的原核表达载体,命名为pET-32a-ompU,将其转化到大肠杆菌E.coli中进行融合表达。37℃,0.5mmol.L-1的IPTG诱导4h,融合蛋白rompU以可溶形式表达,采用Ni-NTA亲和层析法纯化融合蛋白,SDS-PAGE显示纯化后的rompU为与预期大小一致的单一条带。将纯化的rompU免疫SD级大鼠,ELISA检测收获的多克隆抗体血清效价,并通过Western-blot证实该OmpU多克隆抗体可以识别创伤弧菌FJ03-X2株中的天然外膜蛋白OmpU。
- 创伤弧菌 /
- 外膜蛋白OmpU /
- 克隆 /
- 原核表达 /
- 蛋白纯化 /
- 多克隆抗体
Abstract: Vibrio vulnificus is an important pathogen that causes the eel sepsis.In this study,the outer membrane protein U(ompU)gene of Vibrio vulnificus strain FJ03-X2is cloned,and then inserted into the prokaryotic expression vector,named pET32a-ompU.The plasmid was then transformed into E.coli strain to induce the expression of rompU protein.The soluble rompU protein can be expressed under the induction of IPTG(0.5mmol/ L)at 37℃for 4h.Then purified it by Ni-NTA His Binding affinity purification.SDS-PAGE displayed the purified rompU as a single band of the expected size.The purified rompU protein was then injected into SD level rats to produce polyclonal antibody.High specificity antiserum of FJ03-X2rompU is confirmed by ELISA detection.Western-blot confirmed that OmpU polyclonal antibodies can recognize the natural outer membrane protein OmpU in Vibrio vulnificus FJ03-X2.
- Vibrio vulnificus /
- outer membrane protein OmpU /
- cloning /
- prokaryotic expression /
- protein purification /
- polyclonal antibody

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参考文献(15)

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