title: Construction of pTM series plasmids for gene expression in Brucella species.
abstract: 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×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×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.
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.
authors: Mingxing
authors: M
authors: Tian
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.
authors: Jing
authors: J
authors: Qu
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.
authors: Yanqing
authors: Y
authors: Bao
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.
authors: Jianpeng
authors: J
authors: Gao
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.
authors: Jiameng
authors: J
authors: Liu
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.
authors: Shaohui
authors: S
authors: Wang
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.
authors: Yingjie
authors: Y
authors: Sun
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China.
authors: Chan
authors: C
authors: Ding
authors: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Shanghai, China. Electronic address: yus@shvri.ac.cn.
authors: Shengqing
authors: S
authors: Yu
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002002:Brucella / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic
chemical_list: D001426:Bacterial Proteins
keywords: Brucella abortus; High efficiency expression; Inducible expression; Plasmids