Quorum Sensing
The whole-cell cultures are used for the monitoring study. Round-bottom flasks (25 ml). Acidified ethyl acetate: [0.01 % (v/v) ...
pH 7.0 can be obtained by diluting 0.1 M sodium phosphate dibasic and 0.1 M sodium phosphate monobasic. Add 4 ml of 0.1 M sodium ...
Elute PYO, HHQ, and PQS from the SPE cartridge using 0.5 ml of 5 % of 4.5 M ammonium formate in methanol (95 % CH 3 OH:5 % DW, ...
Electrolyte solution for PYO, HHQ, and PQS detection: 50 mM acetate buffer (pH 5.0) with 20 % ACN. Prepare a stock solution of ...
An SPE tube adapter is used to connect the syringe plunger to the SPE sorbent for applying air pressure. Always prepare the aci ...
Govan JR, Deretic V (1996) Microbial patho- genesis in cystic fibrosis: mucoidPseudomonas aeruginosaandBurkholderia cepacia. Mi ...
Chapter 10 Detection and Quantification of Butyrolactones fromStreptomyces Marc Biarnes-Carrera, Rainer Breitling, and Eriko Tak ...
from various non-Streptomycetes [5]. However, even more GBL synthases and repressors have been identified in genome sequence dat ...
2 Materials 2.1 Extraction of Butyrolactones from Solid Culture Trace element solution [17]: 0.1 g/l ZnSO 4 ·7H 2 O, 0.1 g/l Fe ...
10^7 Spores ofS. coelicolorM1152 [18]orS. coelicolorM145 [12]. Sterile water. Razor blades. Ethyl acetate, HPLC grade (seeNote ...
2.3 Detection of Butyrolactones by Means of Antibiotic Bioassay SFM medium [17]: 20 g/l Agar (Difco), 20 g/l mannitol, 20 g/l s ...
Add 10^7 Streptomycesspores/plate, diluted with sterile water in 100 μl total volume, and spread evenly across the entire plate ...
Allow the MgSO 4 to sediment at the bottom of the beaker and then filter. Remove the solvent by rotary evaporation at room temp ...
3.4 Detection of Butyrolactones: Kanamycin Bioassay The kanamycin bioassay is based on the use ofS. coelicolorLW18 or LW94 strai ...
0.25μg/μl solution of SCB1 or 2–3μl of a previously validated methanol extract and let dry for 1 min. Incubate at 30C incubato ...
Fig. 3Kanamycin bioassay results in DNAgar containing 5μg/ml of kanamycin after incubation at 30C for up to 3 days with two dif ...
aliquots of at most 3μl sequentially and let the plates dry in between additions, to avoid excessive spreading of the extract on ...
Wang J, Wang W, Wang L, Zhang G, Fan K, Tan H et al (2011) A novel role of ‘pseudo’γ- butyrolactone receptors in controllingγ-b ...
Part II Methods for the Studying of Quorum Sensing at the Molecular, Physiological, and Population Level ...
Chapter 11 Fluorescence Quenching Studies ofγ-Butyrolactone- Binding Protein (CprB) fromStreptomyces coelicolorA3(2) Jessy Maria ...
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