Quorum Sensing

degradation, and prediction of QQ enzymes from whole genome sequencing data. The discovery of more QQ enzymes will contrib- ute ...

2.2 Further Characterization of QQ Activities of Selected Bacteria Sonicator equipped with a microtip. Water bath. 20 mg/ml pro ...

4.5μg/ml tetracycline and incubate at 28C, 170 rpm for 12 h (seeNote 5). Prepare the A136 X-gal assay solution by inoculating ...

sonication, make sure the microtip is surface sterilized with 70% (vol/vol) ethanol. Sonicate the cell suspension in ice 20 tim ...

tube. Add 2μl of C6-HSL and 20μl of PIPES stock solutions to the tube. Mix gently by pipetting and incubate the mixtures at 28C ...

Sequence the whole genome of Th120 in an available company or institute providing whole genome sequencing service. Gen- erally, ...

Table 1Selected results of BLASTP searching showing four putative QQ enzymes from Th120qseqid sseqid pident length mismatch gapo ...

4 Notes The biosensorA. tumefaciensA136 (pCF218) (pCF372) lacks the Ti plasmid and provides AHL response system, the AHL- respo ...

incubation time might decrease the difference of the normal- izedβ-galactosidase activities between candidate bacteria and negat ...

enzymes. In addition, HPLC-MS should be used to identify the lactonase activity of subsequently expressed proteins [7]. Descrip ...

Greenberg EP, Hastings JW, Ulitzur S (1979) Induction of luciferase synthesis inBeneckea harveyiby other marine bacteria. Arch ...

Chapter 24 Directed Evolution of Quorum-Quenching Enzymes: A Method for the Construction of a Directed Evolution Platform and Ch ...

A thermostable lactonase fromGeobacillus kaustophilus(GKL) was used as the template for the in vitro evolution of enhanced quoru ...

20 min. Cool the molten agar solution to 60C before adding the appropriate amount of antibiotic (100μg/ml ampicillin; 30 μg/ml ...

(b) GKL reverse primer: 5^0 -CCGACCTTACAAGGATCCT- CAAGCCGAGAACAGCGCC-3^0. (c) T7 Pro primer: 5^0 -CGAAATTAATACGACTCACTA- TAGG-3^ ...

55 C for 15 s, and 72C for 30 s, and a final extension of 72 C for 10 min. Prepare a 1% (wt/vol) agarose gel while waiting f ...

Pick five to ten colonies that grew on the selective plate and screen them for the presence of the gene by PCR. Prepare 10μl PC ...

Add 1 ml of the culture grown instep 2in 2-l conical flasks containing 1 l of LB broth supplemented with 100μg/ml ampicillin. L ...

3.4 Construction of a Quorum-Quenching Directed Evolution Platform (Fig.1) 3.4.1 Amplification and Cloning of the luxR- luxCDABE ...

Add 50 ng of the LuxR-LuxCDABE/pUC18R6K-mini- Tn7T-GmR plasmid and 50 ng of the pTNS2 plasmid into 100 μlofE. coliJLD271 electr ...