Testing the effect of coordinate overexpression of the lovK-lovR two component system on the Caulobacter transcriptome
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ABSTRACT: This experiment is testing the effects of coordinate overexpression of the lovK-lovR two-component system on gene expression in Caulobacter crescentus relative to an empty vector control strain. Transcription in two independent cultures of a lovK-lovR overexpression strain (FC438) are compared to two independent cultures of an empty vector control strain (FC423)
Project description:phyR encodes a general stress regulator protein that is conserved among several alpha-proteobacteria. This experiment is quantifying transcription in two Caulobacter strains: one strain is overexpressing phyR (gene CC3477); the other carries a phyR in-frame deletion. Both strains were cultured in M2 defined medium with xylose as the sole carbon source. The first strain (FC626) carries a plasmid integrated into the xylX locus; phyR is fused to the xylX promoter in this plasmid, generating a xylose-inducible phyR overexpression strain. The second strain (FC799) carries an in-frame deletion at the chromosomal phyR locus. Duplicate cultures of: 1) C. crescentus strain CB15 phyR in-frame deletion mutant , and 2) a xylose-inducible phyR overexpression strain, were cultured in M2 xylose defined medium and harvested at OD660=0.3
Project description:SpoT is an Rsh (Rel / Spo homolog) protein, which synthesizes the alarmone ppGpp in response to starvation. This experiment is quantifying transcription in two Caulobacter strains after 5 minutes of glucose starvation: one strain is wild-type CB15N (NA1000); the other strain is NA1000 which has a chromosomal in-frame deletion of the spoT gene. The wild-type strain can synthesize the alarmone ppGpp in response to starvation and the spoT null strain cannot. Both strains were cultured in M2 defined medium with glucose as the sole carbon source, and then washed and incubated in M2 medium lacking glucose for 5 minutes before RNA isolation. Triplicate cultures of: 1) C. crescentus strain CB15N , and 2) CB15N ?SpoT, cultured in M2-glucose media, then starved for glucose for 5 minutes.
Project description:To test the effects of hypoxia on transcription in Caulobacter crescentus, we cultured cells in a New Brunswick bioreactor under controlled conditions. Prior to innoculation, the medium was bubbled with laboratory air at maximum flow and stirred at 300 rpm for 2 hours. After this period, the medium was considered saturated with air and the oxygen probe was set to 100%. Untreated cultures were grown in air-saturated complex medium at 30 degrees C to OD660=0.5 at pH=7 (continuous air-bubbling; 300 rpm stirring). At cell harvest in aerated culture, the dissolved oxygen probe remained above 98%. To subject cells to hypoxia, culture at OD660=0.5, pH=7 was sparged continuously with nitrogen gas; the dissolved oxygen level as measured by the gas probe dropped from 100% to 0% over the course of 5 minutes under this condition. Hypoxic cultures were continually stirred and bubbled with nitrogen for another 20 minutes after the dissolved gas probe read 0%. Hypoxic cells were then harvested for RNA isolation. Four independent biological samples are included in this study. Two batches of cells were subjected to 20 minutes of hypoxia in a bioreactor; two cell batches were highly aerated.
Project description:Comparison in late exponential phase (culture OD600 = 3) of global expression profiles from a Bacillus thuringiensis 407 strain overexpressing transcriptional regulator MogR from xylose inducible vector pHT304-Pxyl, versus an isogenic empty vector control strain, to analyze global expression changes resulting from MogR overexpression
Project description:Comparison of global expression profiles from a Bacillus thuringiensis 407 strain overexpressing the diguanylate cyclase CdgF (BTB_c06420) from the xylose inducible vector pHT304-Pxyl, versus an isogenic empty vector control strain, to analyze global expression changes resulting from CdgF overexpression
Project description:Transcription analysis of M. tuberculosis H37Ra devR overexpressing strain (LIX48) versus empty vector control strain (LIX47); H37Rv devR overexpressing strain t (LIX50) versus empty vector control strain (LIX49) to study the effect of DevR overexpression
Project description:We did transcription profiling on the effect of RCK1 over-expression. rck1 mutant strain was transformed with empty high copy vector pRS425 empty vector), or with RCK1 cloned into pRS425 (the RCK1-overexpressing strain: RCK1 cloned into pRS425).
Project description:M. truncatula was transformed with TT2 (transparent testa2 from Arabidopsis) using Agrobacterium rhizogene and the transgenic hairy roots were used for gene profiling in comparison with empty vector control.