Transcriptional profiling by array of Burkholderia cenocepacia clonal variants IST439 and IST4113
ABSTRACT: 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
Pulmonary colonization of cystic fibrosis (CF) patients with Burkholderia cenocepacia or other bacteria of the Burkholderia cepacia complex (Bcc) is associated with worse prognosis and increased risk of death. During colonization, the bacteria may evolve under the stressing selection pressures exerted in the CF lung, in particular, those resulting from challenges of the host immune defenses, antimicrobial therapy, nutrient availability and oxygen limitation. Understanding the adaptive mechanisms ...[more]
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: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: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 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: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>