Construction and Characterization of a Gradually Inducible Expression Vector for Halobacterium salinarum, Based on the kdp Promoter
Construction and Characterization of a Gradually Inducible Expression Vector for Halobacterium salinarum, Based on the kdp Promoter
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DOI：
10.1128/aem.07155-11
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发表时间：
2012-04-01
影响因子：
4.4
通讯作者：
Greie, Joerg-Christian
中科院分区：
文献类型：
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作者：
Kixmueller, Dorthe;Greie, Joerg-Christian
受生长条件变化控制的逐渐诱导型表达载体是条件致死突变体基因表达的有力工具。此外，受控表达允许监测蛋白质在其表达宿主的各个阶段的过量生产。对于盐生盐杆菌（Halobacterium salinarum），其通常被用作嗜盐古菌的范例，这样的诱导型表达系统迄今不可用。在这里，我们表明，kdp启动子（Pkdp），这有利于基因表达后K+限制，可以用来建立这样一个系统的分子应用。Pkdp具有相当高的表达率，约50倍的增加，可以很容易地通过生长培养基中的K+浓度变化。除了构建表达载体外，我们的工作还描述了表达模式的表征，从而为科学界提供了一个逐渐诱导的表达系统。
Gradually inducible expression vectors which are governed by variations of growth conditions are powerful tools for gene expression of conditionally lethal mutants. Furthermore, controlled expression allows monitoring of overproduction of proteins at various stages in their expressing hosts. For Halobacterium salinarum, which is often used as a paradigm for halophilic archaea, such an inducible expression system is not available to date. Here we show that the kdp promoter (Pkdp), which facilitates gene expression upon K+ limitation, can be used to establish such a system for molecular applications. Pkdp features a rather high expression rate, with an approximately 50-fold increase that can be easily varied by K+ concentrations in the growth medium. Besides the construction of an expression vector, our work describes the characterization of expression patterns and, thus, offers a gradually inducible expression system to the scientific community.