In order to evaluate the influence of 40µM PQS on the global Pseudomonas aeruginosa gene expression, we performed a comparative gene expression analysis of PQS-treated versus untreated P. aeruginosa PAO1 cultures after 5, 11 and 20 h of incubation.
Treatment protocol
P. aeruginosa PAO1 cultures without addition of 40 µM PQS were grown in BHI medium and total RNA was extracted after 11 h of growth
Growth protocol
P. aeruginosa PAO1 cultures without addition of 40 µM PQS were grown in 10 ml BHI medium in a 50 ml flask shaking culture (180 rpm, 37°C) and total RNA was extracted after 11 h of growth
Extracted molecule
total RNA
Label
Biotin-16-ddUTP
Hybridization protocol
Hybridization of the probes was carried out according to the manufacturer’s “Expression Analysis Protocol (p. 25: Modified Fluidic Protocol for P. aeruginosa cDNA Assay)”, see http://www.affymetrix.com:
Description
· Threshold of Signal Log ratio: -1/1; change is I (increased), MI (marginally increased), D (decreased), MD (marginally decreased); change p-value >0.999 or <0.001; only genes that are present (Detection=P) in at least one of the compared chipsets.
