Project description:Halobacterium salinarum NRC-1 was grown in CM media, at 37oC in a waterbath with agitation of 125 rpm under constant light. Analysis of transcriptional changes during growth, in addition to mapping of transcriptome structure under the same conditions, provided interesting insights about regulatory logic within prokaryotic coding regions.
Project description:Halobacterium salinarum NRC-1 was grown in CM media, at 37oC in a waterbath with agitation of 125 rpm under constant light. Analysis of transcriptional changes during growth, in addition to mapping of transcriptome structure under the same conditions, provided interesting insights about regulatory logic within prokaryotic coding regions. Samples were collected at different cell densities, from OD ~0.2 to OD ~5.0. 2 biological replicates were conducted. For each sample, a dye-swap experiment was performed.
Project description:In order to ensure the reproducibility of the transcriptional response of Halobacterium NRC-1 to oxic/anoxic transitions, we repeated global mRNA measurements for the oxygen time series data in GSE5924, except that cultures were equilibrated to low oxygen for 24 hours prior to the start of the experiment rather than 12 hours, and time course sampling continued to 12 hours post-shift to high oxygen rather than stopping at 6 hours. The results of these data suggest that there is good reproducibility between datasets, and that Halobacterium responds robustly to oxic/anoxic transitions. Keywords: time course
Project description:Total RNA hybridizations of Halobacterium salinarum NRC-1 total RNA versus a strain overexpressing TFBd were compared to verify the transcriptome landscape changes. Comparison with TFBd binding sites localized using ChIP-chip data and MeDiChI algorithm (Reiss et al, 2008) allowed observation that TF-binding in the middle of coding sequences can also result in transcription initiation or termination internal to a single annotated protein-coding gene. Total RNA sample of plasmid based expression of Cmyc tagged transcription factor TFBd grown in standard growth media with mevinolin.
