Construction of pTM series plasmids for gene expression in Brucella species

文献类型： 外文期刊

第一作者： Tian, Mingxing

作者： Tian, Mingxing;Qu, Jing;Bao, Yanqing;Gao, Jianpeng;Liu, Jiameng;Wang, Shaohui;Sun, Yingjie;Ding, Chan;Yu, Shengqing

作者机构：

关键词： Brucella abortus;Plasmids;High efficiency expression;Inducible expression

期刊名称：JOURNAL OF MICROBIOLOGICAL METHODS （ 影响因子：2.363； 五年影响因子：2.24 ）

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收录情况： SCI

摘要： Brucellosis, the most common widespread zoonotic disease, is caused by Brucella spp., which are facultative, intracellular, Gram-negative bacteria. With the development of molecular biology techniques, more and more virulence-associated factors have been identified in Brucella spp. A suitable plasmid system is an important tool to study virulence genes in Brucella. In this study, we constructed three constitutive replication plasmids (pTM1-Cm, pTM2-Amp, and pTM3-Km) using the replication origin (rep) region derived from the pBBR1-MCS vector. Also, a DNA fragment containing multiple cloning sites (MCSs) and a terminator sequence derived from the pCold vector were produced for complementation of the deleted genes. Besides pGH-6 x His, a plasmid containing the groE promoter of Brucella spp. was constructed to express exogenous proteins in Brucella with high efficiency. Furthermore, we constructed the inducible expression plasmid pZT-6 x His, containing the tetracycline-inducible promoter pzt1, which can induce expression by the addition of tetracycline in the Brucella culture medium. The constructed pTM series plasmids will play an important role in the functional investigation of Brucella spp. (C) 2016 Elsevier B.V. All rights reserved.

分类号： Q93-3

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