实验基础

Advances in amplification techniques have revolutionized the ability to detect viruses both quantitatively and qualitatively and to study viral load. Real-time polymerase chain reaction (PCR) amplification depends on the ability to detect and quantify a fluorescent reporter ...

Pseudomonas aeruginosa produces a quorum sensing molecule termed the Pseudomonas Quinolone Signal (2-heptyl-3-hydroxy-4-quinolone; PQS) that regulates an array of genes involved in virulence. This chapter addresses four related techniques useful for detecting and quanti ...

The formation of biofilms in conjunction with quorum sensing (QS)-regulated expression of virulence by opportunistic pathogens contributes significantly to immune evasion and tolerance to a variety of antimicrobial treatments. The present protocol describes methods to de ...

Synthetic quorum-sensing systems in mammalian cells has enabled the implementation of time- and distance-dependent bioprocesses, as well as the design of synthetic ecosystems emulating clinically important host–parasite interactions. In this chapter, we provide a detailed ...

Quorum-quenching enzymes are useful as biochemical tools and possible therapeutic proteins. One of the best-characterized families of these catalysts is the N-acyl-l-homoserine lactone (AHL) lactonases, which rely on a dinuclear metal ion active site to hydrolytically cleave the ...

Bacterial quorum sensing (QS) system is a unique target for the development of a new class of drugs that potentially control pathogenicity and attenuate virulence. Thus, it has been of significant interest to discover small organic molecules that modulate QS circuits by competing with the s ...

The exchange of information within and among bacterial populations using small diffusible molecules has been termed “quorum sensing” (QS). Due to the extracellular distribution of the QS autoinducer molecules and the evolutionary highly conserved nature of signaling componen ...

Mammalian paraoxonases (PONs) are a unique, highly conserved family of calcium-dependent esterases consisting of PON1, PON2, and PON3. The PONs can hydrolyze the lactone ring of a range of N-acyl-l-homoserine lactone (AHL) quorum sensing signaling molecules, rendering them inactive. ...

In many pathogenic bacteria, quorum sensing (QS) controls expression of genes that are involved in virulence, production and resistance to antibiotics, formation and maintenance of microbial multicellular consortia on biotic and abiotic surfaces of medical and industrial impo ...

Within environmental communities, there is a constant struggle for survival, as nutrients are often limited. In response, bacteria have developed elaborate methods to deal with competitors. One such mechanism is the coordination of behaviors and function via the exchange of small che ...

The detection of bacterial signaling molecules in liquid or gaseous environments has been occurring in nature for billions of years. More recently, man-made materials and systems has also allowed for the detection of small molecules in liquid or gaseous environments. This chapter will ou ...

The contamination, survival, and possible foodborne disease outbreaks are major issues confronting the food industry. However, from a microbial perspective, any food whether natural or processed is just another environmental niche that is available for colonization. Quorum sen ...

High-performance liquid chromatography (HPLC) coupled in-line with mass spectrometry (MS) permits rapid and specific identification and quantification of N-acyl-l-homoserine lactones (AHLs) and 4-hydroxy-2-alkylquinolines (HAQs). We are presenting here methods for t ...

Autoregulation of genes is often associated with quorum sensing systems where bacteria produce and secrete molecules that allow the cells to communicate with one another, leading to the activation of certain genes at certain population densities. Here we describe the identification ...

Intercellular small molecular weight signaling molecules modulate a variety of biological functions in bacteria. One of the more complex behaviors mediated by intercellular signaling molecules is the suite of activities regulated by quorum sensing molecules. These molecules ...

Microbial pathogens use a wide repertoire of pathogen-associated molecular patterns (PAMPs) that affect host cell responses through activation of intracellular signaling events in a PAMP-specific manner. Here we describe a set of western blot-based methodologies for the evalu ...

In order to study N-acyl homoserine lactone (AHL)-based quorum sensing in vivo, we present a protocol using an Escherichia coli strain equipped with a luxR-based monitor system, which in the presence of exogenous AHL molecules expresses a green fluorescent protein (GFP). Lungs from mice chal ...

Quorum sensing plays a central role in regulating many community-derived symbiotic and pathogenic relationships of bacteria, and as such has attracted much attention in recent years. Acyl-homoserine lactones (AHLs) are important signaling molecules in the quorum sensing gene-r ...

Transcript profiling has improved our understanding of quorum-sensing gene regulation on a global scale. Here, we describe methods for high-quality sample preparation for use in DNA microarray and real-time PCR experiments.

The two-component quorum sensing (QS) system, first described in the marine bacterium Vibrio harveyi and evolutionarily conserved among members of the genus Vibrio, has been best studied in the human pathogen Vibrio cholerae (1, 2). In the V. cholerae QS system, the response to the accumulation ...