Transcription profiling by array of Burkholderia ambifaria during four growth conditions which modulated enacyloxin secretion

ABSTRACT: 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

ORGANISM(S): Burkholderia ambifaria

SUBMITTER: Andrea Sass 

PROVIDER: E-MEXP-2819 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress