An improved tetracycline-inducible expression vector for Staphylococcus aureus
An improved tetracycline-inducible expression vector for Staphylococcus aureus
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DOI：
10.1016/j.plasmid.2008.10.001
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发表时间：
2009-03-01
期刊：
影响因子：
2.6
通讯作者：
Foster, Timothy J.
中科院分区：
文献类型：
--
作者：
Corrigan, Rebecca M.;Foster, Timothy J.
四环素诱导型表达载体 pALC2073 允许克隆的 sasG 基因高水平表达，但未诱导细胞的抑制是有漏洞的。 tetR 启动子的 -10 盒突变为枯草芽孢杆菌共有序列，从而导致 SasG 蛋白表达的完全抑制。 1.28 μ g ml(-1) 脱水四环素的诱导水平与使用 160 ng ml(-1) 四环素的 pALC2073sasG 获得的诱导水平相同，这是可以使用而不抑制细菌生长的最高浓度。因此，pALC2073 的这种变体在未诱导时提供几乎完全的抑制，在诱导时提供高水平的表达。 (C) 2008 Elsevier Inc. 保留所有权利。
The tetracycline-inducible expression vector pALC2073 allowed high level expression of the cloned sasG gene but repression by uninduced cells was leaky. The -10 box of the tetR promoter was mutated to the Bacillus subtitlis consensus, which resulted in complete repression of SasG protein expression. Anhydrotetracycline at 1.28 mu g ml(-1) gave the same high level of induction that was obtained with pALC2073sasG using 160 ng ml(-1) tetracycline, the highest concentration that could be used without inhibiting bacterial growth. This variant of pALC2073 thus offers almost complete repression when uninduced and high levels of expression when induced. (C) 2008 Elsevier Inc. All rights reserved.