Quorum Sensing

2.3 Detection of AQs Using TLC Normal-phase 2020 cm silica 60F254TLC plates. TLC developing tank. Soft top agar: 0.65% (wt/vol ...

in 10 ml of fresh LB or relevant medium. Centrifuge again at 10,000gfor 10 min and discard the supernatant. Repeat the media wa ...

Reconstitute sample extracts in 100μl of methanol and spot 5 μl of each onto the TLC plate. The amount spotted onto the TLC dep ...

3.5 Testing for AQ Production Using a Microtiter Plate Assay Prepare crude bacterial culture supernatants by growing the test b ...

Monitor bioluminescence and OD at 37C using a combined spectrophotometer/luminometer. This measures OD and bio- luminescence f ...

4 Notes Growth ofP. aeruginosacultures for 8 h in LB medium results in the bacteria reaching mid-stationary phase, which is suf ...

Heeb S, Fletcher MP, Chhabra SR, Diggle SP, Williams P, Ca ́mara M (2011) Quinolones: from antibiotics to autoinducers. FEMS Mi ...

Chapter 3 “Hot Stuff”: The Many Uses of a Radiolabel Assay in Detecting Acyl-Homoserine Lactone Quorum-Sensing Signals Amy L. Sc ...

bound by QS signal, it controls gene expression as activator or repressor, depending upon the particular system [1, 2]. Two gene ...

1.2 Techniques to Detect and Identify AHL Compounds A variety of protocols have been developed to detect and quantify AHL signal ...

were first identified, thep-coumaroyl- and isovaleryl-linked AHLs (Fig. 1), they were not detected by the existing repertoire of ...

applied in many ways: to detect novel AHL signals [6, 11]; to detect AHL production in situ for complex environments such as cys ...

3 Methods 3.1 Protocol for AHL Detection During Bacterial Growth This is a general protocol for^14 C-radiolabeling of AHL signal ...

Radiolabel cells by adding 5μCi (~90 nmol,seeNote 1)of [1-^14 C]-methionine (^14 C-met) to the culture for an appropri- ate lab ...

in Figs.3 and 4, the amount of^14 C-AHL produced can vary by ~10,000-fold depending upon the bacterium, labeling time, and exper ...

Dilute the culture (~1:5) in fresh medium. For each experimental comparison condition (e.g., no addition, aro- matic acid mixtu ...

gradient (1 ml/min flow, 70-min profile). Collect 70 1-ml (1-min) fractions in scintillation vials using an automated frac- tion ...

final concentration of 0.1 mM for substrate labeling experiments. Although here we use defined aromatic acid mixtures, one coul ...

Acknowledgements This work was sponsored by the following funding sources: (1) the Agricultural and Food Research Initiative Com ...

Zhu J, Chai Y, Zhong Z, Li S, Winans SC (2003)Agrobacteriumbioassay strain for ultra- sensitive detection of N-acylhomoserine l ...