坏死梭杆菌外膜蛋白基因的克隆与表达

›› 2013, Vol. 40 ›› Issue (6): 57-60.

坏死梭杆菌外膜蛋白基因的克隆与表达

徐晶^1,2, 陈立志², 刘晓颖², 冯二凯², 汪孙杰^1,2, 曹阅²

1. 江苏科技大学生物与化学工程学院, 江苏镇江 212018;
2. 中国农业科学院特产研究所, 吉林省特种经济动物分子生物学省部共建实验室, 吉林长春 130112

收稿日期:2012-11-30 出版日期:2013-06-20 发布日期:2013-06-20
通讯作者: 陈立志 E-mail:tcsclz@126.com
作者简介:徐晶(1986-),女,黑龙江人,硕士,研究方向:分子细菌学。
基金资助:
吉林省科技发展计划(20110223)。

Cloning and Expression of Outer Membrane Proteins (OMP) of Fusobacterium necrophorum (AB)

XU Jing^1,2, CHEN Li-zhi², LIU Xiao-ying², FENG Er-kai², WANG Sun-jie^1,2, CAO Yue²

1. School of Biology and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212018, China;
2. State Key Laboratory for Molecular Biology of Special Economical Animals, Institute of Special Animal and Plant Science, Chinese Academy of Agricultural Sciences, Changchun 130112, China

Received:2012-11-30 Online:2013-06-20 Published:2013-06-20

摘要/Abstract

摘要： 依据GenBank中登录的坏死梭杆菌外膜蛋白(outer membrane protein,OMP)基因序列,设计1对引物进行PCR扩增OMP片段并克隆至pET-28a中,构建原核表达重组质粒pET-28a-OMP。将pET-28a-OMP转化到大肠杆菌BL21(DE3)感受态中,表达含2个His的重组蛋白。SDS-PAGE和Western blotting分析结果表明,该重组蛋白分子质量为93 ku,为包涵体表达,具有反应原性。本试验结果为坏死梭杆菌OMP免疫机制的研究提供了基础数据。

关键词: 坏死梭杆菌; OMP; 诱导表达; 免疫活性

Abstract: The outer membrane protein (OMP) was deposited in GenBank. The upstream and downstream primers were designed, and OMP gene was amplified by PCR. Then the target gene was inserted into the vector pMD18-T Simple and pET-28a, and the recombinant vectors were used to select and be transformed into E.coli TOP10 and BL21 (DE3), respectively. The plasmid from E.coli TOP10 and BL21 (DE3) was digested by restricted endonuclease enzyme of XhoⅠand EcoRⅠ. Then the plasmid of enzyme digestion was analyzed by its sequence. The recombinant protein was expressed by IPTG induction. We successfully obtained OMP by identification of SDS-PAGE, and result of Western blotting showed that the OMP had immune activity.

Key words: Fusobacterium necrophorum; outer membrane protein (OMP); induction expression; immune activity

中图分类号:

Q786

徐晶, 陈立志, 刘晓颖, 冯二凯, 汪孙杰, 曹阅. 坏死梭杆菌外膜蛋白基因的克隆与表达[J]. , 2013, 40(6): 57-60.

XU Jing, CHEN Li-zhi, LIU Xiao-ying, FENG Er-kai, WANG Sun-jie, CAO Yue. Cloning and Expression of Outer Membrane Proteins (OMP) of Fusobacterium necrophorum (AB)[J]. , 2013, 40(6): 57-60.

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