Qualitative and quantitative determination of quorum sensing inhibition in vitro
Research output: Contribution to journal › Journal article › peer-review
Standard
Qualitative and quantitative determination of quorum sensing inhibition in vitro. / Jakobsen, Tim Holm; van Gennip, Maria; Christensen, Louise Dahl; Bjarnsholt, Thomas; Givskov, Michael.
In: Methods in Molecular Biology, Vol. 692, No. 3, 01.01.2011, p. 253-263.Research output: Contribution to journal › Journal article › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Qualitative and quantitative determination of quorum sensing inhibition in vitro
AU - Jakobsen, Tim Holm
AU - van Gennip, Maria
AU - Christensen, Louise Dahl
AU - Bjarnsholt, Thomas
AU - Givskov, Michael
PY - 2011/1/1
Y1 - 2011/1/1
N2 - The formation of biofilms in conjunction with quorum sensing (QS)-regulated expression of virulence by opportunistic pathogens contributes significantly to immune evasion and tolerance to a variety of antimicrobial treatments. The present protocol describes methods to determine the in vitro efficacy of potential quorum sensing inhibitors (QSIs). Work on Pseudomonas aeruginosa has shown that chemical blockage of QS is a promising new antimicrobial strategy. Several live bacterial reporter systems been developed to screen extracts and pure compounds for QSI activity. Here we describe the usage of reporter strains consisting of a lasB-gfp or rhlA-gfp fusion in P. aeruginosa for qualitative and quantitative evaluation of the inhibition of the two major QS pathways, monitored as reduced expression of green fluorescence. By the use of an in vitro flow cell system it is possible to study the QSI activity by monitoring its ability to interfere with the protective functions of bacterial biofilm. For evaluation of the global effects of QSI compounds, we present a protocol for the DNA microarray-based transcriptomics. Using these in vitro methods it is possible to evaluate the potential of various QSI compounds.
AB - The formation of biofilms in conjunction with quorum sensing (QS)-regulated expression of virulence by opportunistic pathogens contributes significantly to immune evasion and tolerance to a variety of antimicrobial treatments. The present protocol describes methods to determine the in vitro efficacy of potential quorum sensing inhibitors (QSIs). Work on Pseudomonas aeruginosa has shown that chemical blockage of QS is a promising new antimicrobial strategy. Several live bacterial reporter systems been developed to screen extracts and pure compounds for QSI activity. Here we describe the usage of reporter strains consisting of a lasB-gfp or rhlA-gfp fusion in P. aeruginosa for qualitative and quantitative evaluation of the inhibition of the two major QS pathways, monitored as reduced expression of green fluorescence. By the use of an in vitro flow cell system it is possible to study the QSI activity by monitoring its ability to interfere with the protective functions of bacterial biofilm. For evaluation of the global effects of QSI compounds, we present a protocol for the DNA microarray-based transcriptomics. Using these in vitro methods it is possible to evaluate the potential of various QSI compounds.
U2 - 10.1007/978-1-60761-971-0_18
DO - 10.1007/978-1-60761-971-0_18
M3 - Journal article
C2 - 21031317
VL - 692
SP - 253
EP - 263
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
IS - 3
ER -
ID: 32470212