2 - BACTERIOWEB

How to characterize efflux mechanisms
Plasmid or transposon encoded efflux systems
– Multiresistance phenotype
– Detection of efflux gene(s): PCR, nucleic probes
Upregulation of intrinsic efflux systems
– Protein levels
♦ Western blotting of membrane extracts with specific antibodies
– mRNA levels
♦ Northern blot, MacroArray, MicroArray
♦ Real Time RT-PCR (Light Cycler, Taq Man, I Cycler…)
– Intracellular accumulation of antibiotics
♦ [ 3 H] ou [ 14 C] radiolabeled or fluorescent compounds (BET,
acriflavine…)
– Sequencing of regulatory genes
40