Burkholderia cenocepacia is a significant opportunistic pathogen in individuals with cystic fibrosis. ShvR, a LysR-type transcriptional regulator, has previously been shown to influence colony morphology, biofilm formation, virulence in plant and animal infection models, and some quorum-sensing-dependent phenotypes. In the present study, it was shown that ShvR negatively regulates its own expression, as is typical for LysR-type regulators. The production of quorum-sensing signal molecules was de ...[more]
Project description:In this work the genomic expression of two Burkholderia cenocepacia clonal variants (IST439 and IST4113) were compared. These isolates were collected from a chronically colonized cystic-fibrosis patient that has died from the cepacia syndrome. IST439 was the first B. cenocepacia isolate recovered from the patient and it is thought to have initiated the infection while the IST4113 isolate was recovered three years later, after a period of exacerbated infection that compelled the patient to hospitalization and intravenous therapy with gentamicine and ceftazidime. Among other phenotypic differences, IST4113 is much more resistant to a wide range of antimicrobials tested with very different biological targets
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>
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>