Reproducibility M-Tram of Enterococcus faecium strain E1163
ABSTRACT: Two independent aliquots containing approximately 10^7 cells from the transposon mutant of E. faecium E1162 were grown at 37 C for 20 hours in 20 ml of BHI broth. Genomic DNA was isolated from the two replicate cultures and used for the generation of cDNA. The cDNA samples were labeled with Cy3 and Cy5 respectively and hybridized to a microarray that was designed using the E. faecium E1162 genome sequence.
Enterococcus faecium has become a nosocomial pathogen of major importance, causing infections that are difficult to treat owing to its multi-drug resistance. In particular, resistance to the β-lactam antibiotic ampicillin has become ubiquitous among clinical isolates. Mutations in the low-affinity penicillin binding protein PBP5 have previously been shown to be important for ampicillin resistance in E. faecium, but the existence of additional resistance determinants has been suggested. Here, we ...[more]
Project description:Enterococcus faecium is an important opportunistic pathogen emerging in hospitals worldwide. We identified a new MarR family global regulator in E. faecium, named AsrR (antibiotic and stress response regulator). We phenotipically characterized key role for AsrR in E. faecium pathogenicity. The aim of the microarray-based experiments was to investigate the AsrR regulon in E. faecium. We constructed a mutant strain deleted for the asrR gene, we complemented the mutant and, finally, we observed genes expression in these strains in comparison with the wild-type strain. (The parental HM1070 was used to delete the asrR gene and to obtain the mutant strain. Then, the mutant strain was used to restore the asrR gene and to obtain the complemented strain.) This approach will allow us to identify the genes regulated by AsR to clarify its role in E. faecium.
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