Lee S H, Butler S M, Camilli A
Tufts University School of Medicine, Department of Molecular Biology and Microbiology, 136 Harrison Avenue, Boston, MA 02111, USA.
Proc Natl Acad Sci U S A. 2001 Jun 5;98(12):6889-94. doi: 10.1073/pnas.111581598.
We devised a noninvasive genetic selection strategy to identify positive regulators of bacterial virulence genes during actual infection of an intact animal host. This strategy combines random mutagenesis with a switch-like reporter of transcription that confers antibiotic resistance in the off state and sensitivity in the on state. Application of this technology to the human intestinal pathogen Vibrio cholerae identified several regulators of cholera toxin and a central virulence gene regulator that are operative during infection. These regulators function in chemotaxis, signaling pathways, transport across the cell envelope, biosynthesis, and adherence. We show that phenotypes that appear genetically independent in cell culture become interrelated in the host milieu.
我们设计了一种非侵入性基因筛选策略，以在完整动物宿主的实际感染过程中鉴定细菌毒力基因的正调控因子。该策略将随机诱变与转录的开关式报告基因相结合，该报告基因在关闭状态下赋予抗生素抗性，在开启状态下赋予敏感性。将该技术应用于人类肠道病原体霍乱弧菌，鉴定出了几种霍乱毒素调节因子以及一种在感染过程中起作用的核心毒力基因调节因子。这些调节因子在趋化作用、信号通路、跨细胞膜运输、生物合成和黏附中发挥作用。我们表明，在细胞培养中看似遗传独立的表型在宿主环境中变得相互关联。
