3 with primers PL372 and PL373. SAHA mouse EB 1.3 MG1655 rpoS::Tn10-tet [33] Plasmids and phage Relevant characteristics Reference pBAD24 AmpR, ColE1 [70] pBAD24-Δ1 pBAD24 derivative with a modified polylinker; carries an unique NcoI site overlapping the araBp transcription start this

work pBADpnp pBAD24 derivative; harbours an EcoRI-HindIII fragment of pEJ01 that carries the pnp gene this work pBADrnb pBAD24 derivative; harbours an HindIII-XbaI fragment of pFCT6.9 that carries the rnb gene this work pBADrnr pBAD24-Δ1 derivative; harbours the rnr gene (obtained by PCR on MG1655 DNA with FG2474-FG2475 oligonucleotides) between NcoI-HindIII sites this work pΔLpga pJAMA8 derivative, harbours the -116 to +32 region relative to the pgaABCD transcription start site cloned into the SphI/XbaI sites this work pEJ01 carries a His-tagged pnp allele [71] pFCT6.9 carries a His-tagged rnb allele [72]; received from Cecilia Arraiano pGZ119HE oriVColD; CamR [73] pJAMA8 AmpR, ColE1; luxAB based promoter-probe vector. [37] pLpga1 pJAMA8 derivative, harbours the -116 to +234 region relative to the pgaABCD transcription start site cloned into the SphI/XbaI sites. this work pLpga2 pJAMA8 derivative, harbours a translational

fusion of pgaA promoter, regulatory QNZ mouse region and first 5 codons of pgaA (-116 to +249 relative to transcription start site) with luxA ORF (Open Reading Frame). this work pTLUX pJAMA8 derivative, harbours

ptac promoter of pGZ119HE cloned into the SphI/XbaI sites. this work P1 HTF High transduction frequency phage P1 derivative [74]; received from Richard Calendar Cell aggregation and adhesion assays Cell aggregation was assessed as follows: overnight www.selleckchem.com/products/epoxomicin-bu-4061t.html cultures grown in LD at 37°C on a rotatory device were diluted 50-fold in 50 ml of M9Glu/sup in a 250 ml flask. The cultures were then incubated at 37°C with shaking at 100 rpm. Cell adhesion to the flask walls was assessed in overnight cultures grown Silibinin in M9Glu/sup medium at 37°C. Liquid cultures were removed and cell aggregates attached to the flask glass walls were stained with crystal violet for 5 minutes to allow for better visualization. Quantitative determination of surface attachment to polystyrene microtiter wells was carried out using crystal violet staining as previously described [33]. Binding to Congo red (CR) was assessed in CR agar medium (1% casamino acid, 0.15% yeast extract, 0.005% MgSO4, 2% agar; after autoclaving, 0.004% Congo red and 0.002% Coomassie blue). Overnight cultures in microtiter wells were replica plated on CR agar plates, grown for 24 h at 30°C, and further incubated 24 h at 4°C for better detection of staining. Gene expression determination RNA extraction, Northern blot analysis and synthesis of radiolabelled riboprobes by in vitro transcription with T7 RNA polymerase were previously described [34, 35].