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SRX2918108: GSM2668035: Fermentor N°1 Sample 8h (D3); Clostridium beijerinckii; RNA-Seq
1 ILLUMINA (NextSeq 500) run: 11.1M spots, 564.2M bases, 358.9Mb downloads

Submitted by: NCBI (GEO)
Study: RNA-seq analysis of glucose fermentation by the natural Isopropanol producer Clostridium beijerinckii DSM6423
show Abstracthide Abstract
Purpose:C. beijerinckii DSM 6423 is the most cited natural IpOH producer. Improving the natural production of this strain through a targeted approach required a full sequencing and characterization of its genome, together with transcriptomic analyses of its own regulations.The goals of this study are then to evaluate the transcriptional profile (RNA-Seq) of C. beijerinckii DSM6423, a natural isopropanol producer, during a fermentation of glucose in controlled bioreactors. Methods : A RNA-Seq approach was chosen in order to have a timelapse study of DSM 6423 throughout the fermentation process. Three independent duplicate fermentations of DSM 6423 were carried out in bioreactors on three different weeks, showing good reproducibility. On each cultivation, five biomass samples were collected for RNA-Seq analyses.and DNA was eliminated after DNAse I treatment (AM1906, Invitrogen). The 15 resulting RNA samples were sequenced and analyzed using the previously reconstructed genome of DSM 6423. Results: Using a data analysis workflow (TAMARA) developed by the Genoscope platform of Evry, we were able to highlight the transcriptional regulation along the fermentation by calculating the transcription profiles of each gene, using the 3h sample as reference. Clustering was performed using CAST algorithm revealed 8 clusters containing 953 genes and corresponding to genes up-regulated at 6, 8, 11 or 24 hours and gene down-regulated at 6, 8, 11 or 24 hours. Conclusion : Such analyses were carried out in this study and provide useful data to better understand the genetic background and the physiological specificities of C. beijerinckii DSM6423 isopropanol producer. Notably, this work is the first omic study of a natural IBE producer. The data gathered needs time for proper exploitation, but a better understanding of the metabolic pathways and various genes involved opens the door for future targeted approaches. Overall design: Comparative analysis of samples taken from a serie a fermentations performed in controled conditions
Sample: Fermentor N°1 Sample 8h (D3)
SAMN07236928 • SRS2284151 • All experiments • All runs
Library:
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Cell lysis and RNA isolation was done with the TRIzol Plus RNA purification kit (AMbion). 5 mL TRIzol reagent is added to the frozen pellet followed by 1 mL chloroform after cell lysis. After shacking and centrifugation, the upper phase is mixed with an equal volume of 70% ethanol before transfer to PureLink RNA Spin Cartridges furnished with the kit. Kit instructions allow to obtain purified total RNA which is further processed by cleanup with the RNeasy mini kit (Qiagen). DNA is eliminated after DNAse I treatment (AM1906, Invitrogen). 20µL RNA is treated with 2 µL DNAse in 50 µL nuclease free water for 30 min at 37°C before addition of another 1.5 µL DNAse and 30 min extra incubation. Treatment is done twice and is followed by RNA cleanup with the RNeasy kit. A full quality assessment of the RNA, quantification using Nanodrop, PCR to check for residual DNA and migration on 1% agarose gel is done before depletion of ribosomal RNA with MicrobExpress kit (ThermoFischer). Depletion uses 2-10 µg total RNA in 15 µL water. RNA libraries were prepared for sequencing using standard Illumina protocols
Experiment attributes:
GEO Accession: GSM2668035
Links:
Runs: 1 run, 11.1M spots, 564.2M bases, 358.9Mb
Run# of Spots# of BasesSizePublished
SRR568294911,062,034564.2M358.9Mb2018-03-05

ID:
4171394

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