Howard Hughes Medical Institute and Department of Molecular Microbiology and Microbiology, Tufts University School of Medicine, 150 Harrison Avenue, Boston, MA 02111, USA.
Science. 2012 Dec 14;338(6113):1440-4. doi: 10.1126/science.1229556.
Interactions between hosts and pathogens are complex, so understanding the events that govern these interactions requires the analysis of molecular mechanisms operating in both organisms. Many pathogens use multiple strategies to target a single event in the disease process, confounding the identification of the important determinants of virulence. We developed a genetic screening strategy called insertional mutagenesis and depletion (iMAD) that combines bacterial mutagenesis and RNA interference, to systematically dissect the interplay between a pathogen and its host. We used this technique to resolve the network of proteins secreted by the bacterium Legionella pneumophila to promote intracellular growth, a critical determinant of pathogenicity of this organism. This strategy is broadly applicable, allowing the dissection of any interface between two organisms involving numerous interactions.
宿主与病原体之间的相互作用非常复杂，因此要理解控制这些相互作用的事件，就需要分析在两个生物体中起作用的分子机制。许多病原体采用多种策略来针对疾病过程中的单一事件，从而混淆了对毒力的重要决定因素的识别。我们开发了一种称为插入诱变和耗尽（iMAD）的遗传筛选策略，该策略结合了细菌诱变和 RNA 干扰，以系统地剖析病原体与其宿主之间的相互作用。我们使用该技术解析了促进细菌军团菌细胞内生长的细菌分泌蛋白网络，这是该生物体致病性的关键决定因素。该策略具有广泛的适用性，可用于剖析涉及众多相互作用的两个生物体之间的任何界面。
