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一种方便、稳健的活体报告基因系统,可用于监测人类病原体幽门螺杆菌中的基因表达。

A convenient and robust in vivo reporter system to monitor gene expression in the human pathogen Helicobacter pylori.

机构信息

Department of Biology, University of Bologna, Bologna, Italy.

出版信息

Appl Environ Microbiol. 2012 Sep;78(18):6524-33. doi: 10.1128/AEM.01252-12. Epub 2012 Jul 6.

Abstract

Thirty years of intensive research have significantly contributed to our understanding of Helicobacter pylori biology and pathogenesis. However, the lack of convenient genetic tools, in particular the limited effectiveness of available reporter systems, has notably limited the toolbox for fundamental and applied studies. Here, we report the construction of a bioluminescent H. pylori reporter system based on the Photorhabdus luminescens luxCDABE cassette. The system is constituted of a promoterless lux acceptor strain in which promoters and sequences of interest can be conveniently introduced by double homologous recombination of a suicide transformation vector. We validate the robustness of this new lux reporter system in noninvasive in vivo monitoring of dynamic transcriptional responses of inducible as well as repressible promoters and demonstrate its suitability for the implementation of genetic screens in H. pylori.

摘要

三十年来的深入研究极大地促进了我们对幽门螺杆菌生物学和发病机制的理解。然而,缺乏方便的遗传工具,特别是可用报告系统的效果有限,显著限制了基础和应用研究的工具包。在这里,我们报告了基于 Photorhabdus luminescens luxCDABE 盒构建的生物发光幽门螺杆菌报告系统。该系统由一个无启动子的 lux 受体菌株组成,其中启动子和感兴趣的序列可以通过自杀转化载体的双同源重组方便地引入。我们验证了这个新的 lux 报告系统在非侵入性体内监测诱导和可诱导启动子的动态转录反应中的稳健性,并证明了它在幽门螺杆菌中实施遗传筛选的适用性。

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