Construction of plasmid-based expression vectors for Bacillus subtilis exhibiting full structural stability
Construction of plasmid-based expression vectors for Bacillus subtilis exhibiting full structural stability
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DOI：
10.1016/j.plasmid.2005.05.001
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发表时间：
2005-11-01
期刊：
影响因子：
2.6
通讯作者：
Schumann, W
中科院分区：
文献类型：
--
作者：
Nguyen, HD;Nguyen, QA;Schumann, W
已经构建了一系列基于质粒的表达载体，其允许重组蛋白在枯草芽孢杆菌菌株中的稳定细胞内表达。这些表达载体基于最近描述的大肠杆菌-B。枯草杆菌穿梭载体pMTLBS 72，其以θ环形式复制。除了弱组成型启动子P-lepA外，我们还插入了三种不同的可控启动子：P-gsiB，其可以通过热和酸休克诱导，以及通过乙醇诱导，P-xylA和P-spac，其分别响应于木糖和IPTG的加入。这些表达载体的多功能性通过将它们的启动子融合到报告基因和通过用其中三种过表达HtpG蛋白来证明。所有重组载体均表现出完全的结构稳定性。(c)2005年爱思唯尔公司All rights reserved.
A series of plasmid-based expression vectors have been constructed allowing stable intracellular expression of recombinant proteins in Bacillus subtilis strains. These expression vectors are based on the recently described Escherichia coli-B. subtilis shuttle vector pMTLBS72 which replicates as theta circles. Besides the weak constitutive promoter P-lepA, we inserted three different controllable promoters: P-gsiB, which can be induced by heat and acid shock, and by ethanol, P-xylA and P-spac which respond to the addition of xylose and IPTG, respectively. The versatility of these expression vectors was demonstrated by fusing their promoters to a reporter gene and by overexpression of the HtpG protein with three of them. All recombinant vectors exhibited full structural stability. (c) 2005 Elsevier Inc. All rights reserved.