Project description:The effects of temperature stress on transcriptome of purple non sulfur bacterium R. capsulatus were investigated by comparing expression profiles under optimum hydrogen production condition (30°C), heat (42°C) and cold (4°C) stress conditions.
Project description:The effects of temperature stress on transcriptome of purple non sulfur bacterium R. capsulatus were investigated by comparing expression profiles under optimum hydrogen production condition (30M-BM-0C), heat (42M-BM-0C) and cold (4M-BM-0C) stress conditions. Bacteria were grown anaerobically under continuous illumination (200 W/m2) in 150 ml volume photobioreactors at 30M-BM-0C for 48 hours. Then cold stress (4M-BM-0C) and heat stress (42M-BM-0C) were applied in parallel. For microarray analysis, RNA samples from 1.5x109 cells were collected after 2 hours and 6 hours of stress applications. A microarray chip for Rhodobacter capsulatus was not commercially available. For that reason, a GeneChipM-BM-. array was custom designed and manufactured by Affymetrix, Santa Clara, California. The nucleotide sequence of the bacterium is available in GenBank with the accession numbers CP001312 for the chromosome and CP001313 for the plasmid pRCB133. The feature size of the antisense DNA array was 11 micron and the format was 100-3660. A total of 4,052 probe sets for open reading frames (ORFs) and 200 intergenic sequences with longer than 300bp were placed on the microarray. The cDNA synthesis, labeling, and hybridization protocols were performed according to the Affymetrix Expression Analysis Technical Manual given for prokaryotic samples
Project description:Global transcriptome analyses at growth before and after 10 min of photooxidative stress were applied to monitor stress dependent gene expression in the alpha-proteobacterium Rhodobacter capsulatus. Transcriptome profiles of pigmented cultures with high aeration were monitored before and after the onset of singlet oxygen stress.
Project description:Transcriptomic data has been previously used to determine expression patterns in various R. capsulatus strains using custom made Affymetrix microarrays. Additional expression analyses have since been carried out to obtain preliminary data on certain wild type and mutant strains that have not been reported on. We used all current microarray expression data available and constructed an R. capsulatus gene co-expression network and performed functional analysis of identified gene modules. RNA was harvested from various wild type and mutant R. capsulatus strains grown under photoheterotrophic conditions to different growth phases. The samples were hybridized to custom made R. capsulatus Affymetrix microarrays according to manufacturers recommendations. The raw data was RMA normalized and log transformed.
Project description:Transcriptomic data has been previously used to determine expression patterns in various R. capsulatus strains using custom made Affymetrix microarrays. Additional expression analyses have since been carried out to obtain preliminary data on certain wild type and mutant strains that have not been reported on. We used all current microarray expression data available and constructed an R. capsulatus gene co-expression network and performed functional analysis of identified gene modules.
Project description:Gene Co-Expression Network Analysis in Rhodobacter capsulatus and Application to Comparative Expression Analysis of Rhodobacter sphaeroides
Project description:In this study, we achieved a global view of Cu-responsive changes in the prokaryotic model organism Rhodobacter capsulatus using label-free quantitative differential proteomics. Semi-aerobically grown cells under heterotrophic conditions in minimal medium (~ 0.3 M Cu, optimal for growth) were compared with cells grown similarly but supplemented with either 5 M Cu or with 5 mM of the Cu-chelator bathocuproine sulfonate. Mass spectrometry based bottom-up proteome analyses of unfractionated cell lysates identified with high confidence 2430 of the 3632 putative proteins encoded by the genome.
