Transcription profiling by array of Burkholderia cenocepacia J2315 grown to mid-stationary phase in basal salt medium with glucose or glycerol substrates
ABSTRACT: B. cenocepacia J2315 was grown to mid-stationary phase in basal salt medium with two different substrates: 20 mM glucose or 40 mM glycerol. Cells were harvested after 30 hours incubation at 37 degrees centigrade. The expression profile was compared to cells grown on the same medium and harvested in mid-log phase.
Project description:B. cenocepacia J2315 was grown to mid-log phase in different media: LB broth, iso-sensitest broth, basal salt medium with glucose<br>at pH 7 and pH 6, basal salt medium at pH 7 with a reduced iron content, and basal salt medium with glycerol
Project description:Gene expression was examined during four growth conditions which modulated antimicrobial secretion in B. ambifaria AMMD. A minimal salts medium (BSM; Hareland et al. 1975. J Bacteriol 121:272-85) with 0.05% yeast extract was used for all comparisons with the carbon source (4 g/L) and growth condition varied as follows: (i) liquid culture with glucose; (ii) liquid culture with glycerol; (iii) growth on agar with glucose, and (iv) growth on agar with glycerol. For the liquid cultures, 25 ml of growth medium was placed a 250 ml conical flask and inoculated with 0.5 ml of a starter culture of strain AMMD which had been optically adjusted to O.D. 1 at 600 nm and corresponded to a viable count of 10 million colony forming units per ml. After growth with shaking (150 rpm) for 30 h at 30 degrees Celcius, the O.D. 600 nm of the cultures was measured to enable an estimation of cell density to be made, they were then snap-chilled and harvested as previously described (Drevinek et al. 2008. BMC Infect Dis 8:121), and frozen at -80 degrees Celcius. Growth on agar was performed by laying a sterile 47 mm nitrocellulose filter (0.22 ?m) onto a plate of the BSM agar and the same starter culture of AMMD as used for the liquid growth spread over a 2 cm diameter circular area using a sterile cotton swab. The agar cultures were incubated for 30 h at 30 degrees Celcius, the filters removed to sterile 50 ml conical tubes and the bacteria washed off by gentle pipeting using 2 ml ice cold BSM (without any carbon source or yeast extract). This resuspension of surface grown bacteria was processed prior to RNA extraction in an exactly the same way as the liquid cultures
Project description:The response of antibiotic adapted resistant mutants of B. cenocepacia J2315 to antibiotic stress was investigated using expression profiling of three biological replicates and comparing the profiles to the J2315 parent control grown without antibiotics.<br>A reference design was used with Cy3 labeled genomic DNA of B. cenocepacia J2315 as common reference. Three test conditions with three biological replicates each were compared to three replicates of the control condition.<br>Test conditions: J2315-A grown in the presence of 250 ug per ml amikacin, J2315-M grown in the presence of 8 ug per ml meropenem and J2315-T grown in the presence of 60 ug per ml trimethoprim and 300 ug per ml sulfamethoxazole.<br>Control condition: J2315 parent strain grown without antibiotics.
Project description:Global gene expression in three clinical isolates of the ET12 lineage of Burkholderia cenocepacia was compared by microarray analysis: Strain J2315 (isolated 1989), and strains BCC1616 (HI4277) and BCC1617 (HI4283), both isolated in 2008.
Project description:B. cenocepacia J2315 was grown on LB medium to mid-stationary phase at OD 0.3 under full aeration and then transferred into a 50 ml centrifuge tube and placed upright into a CampyGen Compact (Oxoid, Basingstoke, UK) plastic pouch containing the gas generating paper sachet. The pouch was sealed immediately and then the cap of the centrifuge tube loosened to allow exchange of atmosphere between centrifuge tube and pouch. <br>The culture was further incubated at 37 degrees centigrade and 150 rpm for 2-3 hours until it reached OD 0.5. The centrifuge tube was sealed before opening the pouch and then snap-cooled before harvest to minimise exposure to atmospheric oxygen and to ensure expression profiles reflect the lower oxygen concentration.<br>The expression profile was compared to cells grown to OD 0.5 at atmospheric oxygen concentrations.<br>
Project description:B. cenocepacia J2315 was grown on LB medium to mid-stationary phase at O.D. 0.5 at 150 rpm in a shaking incubator at two different temperatures: 37 degrees and 20 degrees centigrade.
Project description:Burkholderia cenocepacia J2315 was grown at 37 degrees centigrade in LB broth to the beginning of stationary phase (18 hours incubation time). <br>The expression profile was compared to cultures harvested in mid-log phase (7 hours incubation time).<br>