Rice Scott A, McDougald Diane, Givskov Michael, Kjelleberg Staffan
Centre for Marine Bio-Innovation, and The School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, Australia.
Methods Mol Biol. 2008;431:55-68. doi: 10.1007/978-1-60327-032-8_5.
Bacteria communicate with other members of their community through the secretion and perception of small chemical cues or signals. The recognition of a signal normally leads to the expression of a large suite of genes, which in some bacteria are involved in the regulation of virulence factors, and as a result, these signaling compounds are key regulatory factors in many disease processes. Thus, it is of interest when studying pathogens to understand the mechanisms used to control the expression of virulence genes so that strategies might be devised for the control of those pathogens. Clearly, the ability to interfere with this process of signaling represents a novel approach for the treatment of bacterial infections. There is a broad range of compounds that bacteria can use for signaling purposes, including fatty acids, peptides, N-acylated homoserine lactones, and the signals collectively called autoinducer 2 (AI-2). This chapter will focus on the latter two signaling systems as they are present in a range of medically relevant bacteria, and here we describe assays for determining whether an organism produces a particular signal and assays that can be used to identify inhibitors of the signaling cascade. Lastly, the signal detection and inhibition assays will be directly linked to the expression of virulence factors of specific pathogens.
细菌通过分泌和感知小的化学信号与群落中的其他成员进行交流。信号的识别通常会导致大量基因的表达,在一些细菌中,这些基因参与毒力因子的调控,因此,这些信号化合物是许多疾病过程中的关键调控因子。因此,在研究病原体时,了解用于控制毒力基因表达的机制是很有意义的,这样就可以设计出控制这些病原体的策略。显然,干扰这种信号传导过程的能力代表了一种治疗细菌感染的新方法。细菌可用于信号传导目的的化合物种类繁多,包括脂肪酸、肽、N-酰基高丝氨酸内酯,以及统称为自诱导物2(AI-2)的信号。本章将重点关注后两种信号系统,因为它们存在于一系列与医学相关的细菌中,在此我们描述用于确定生物体是否产生特定信号的测定方法,以及可用于识别信号级联抑制剂的测定方法。最后,信号检测和抑制测定将直接与特定病原体毒力因子的表达相关联。