Quorum Sensing

Centrifuge the lysate at 10,000gat 4C for 30 min. Take and save the cleared supernatant: it represents the soluble fraction. ...

independent transposition events and not siblings. As a con- trol, and to check the quality of the plates that will be used the ...

4 Notes Fusions can be generated in two main ways. One method is to clone fragments of DNA into a plasmid vector that contains ...

to be encoded by the same genome, it is not appropriate to refer to these LuxR proteins as LuxR solos. Instead, they are non-AHL ...

and rearrange quite frequently. Find a new source of the vector. Another issue is that the RP4 conjugative apparatus must be abl ...

Fuqua C (2006) The QscR quorum-sensing regulon ofPseudomonas aeruginosa: an orphan claims its identity. J Bacteriol 188:3169–31 ...

ClustalW and ClustalX. Curr Protoc Bioinfor- matics Chapter 2:Unit 2 3 Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: imp ...

Chapter 13 Enzymatic Assays to Investigate Acyl-Homoserine Lactone Autoinducer Synthases Daniel Shin and Rajesh Nagarajan Abstra ...

this class of enzymes such as (1) the substrates for AHL synthase have low turnover (kcat) and high nonenzymatic background rate ...

and substituted acyl-chain (EsaI:3-oxo-C6-ACP, BmaI3:3- hydroxy-C8-ACP) [8–16]. Despite the diversity in the type of AHL synthas ...

during acylation (Fig.3); (3) a nucleosidase-xanthine oxidase-cou- pled assay to determine the amount of MTA released upon lacto ...

2.2 Reagents and Equipment 300–500μM Dichlorophenolindophenol (DCPIP) stock solu- tion (10) in nanopure water. 3–10 mM SAM sto ...

110 a b c 100 90 80 70 60 50 40 30 20 10 5.0 5.1 holo-ACP C4-ACP SAM MTA 5.2 5.3 5.4 5.5 5.6 Minutes mAU 1400 1200 1000 800 600 ...

3 Methods 3.1 Dichloro-pheno- lindophenol (DCPIP) Colorimetric Assay 3.1.1 Preparation of Stock Solutions Prepare the following ...

Determine the time it takes for the change inA 600 to level off. This should be the minimum incubation time necessary to reduce ...

Convert the slope in Abs/s toμM/min as mentioned in Subheading3.1.2. Subtract the background rate from this rate to calculate t ...

3.3 Methylthioa- denosine Nucleosidase- Xanthine Oxidase- Coupled Spectrophotometric Assay 3.3.1 Determination of Background Rat ...

3.4 HPLC Assay 3.4.1 Column Wash Run the column clean method (Table4). After the column wash, equilibrate the column for 3–5 mi ...

3.4.2 Standard Curve for Lactonization Half- Reaction Analytes Prepare 1 mM MTA stock solution in nanopure water (seeNote 11 ). ...

3.4.4 Quench Test 1. Add 170μl water, 30μl 1 M HEPES buffer pH 7.3 (10), 30 μl10 acyl-ACP/acyl-CoA stock, 30 μl10 AHL synthas ...