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SRX24095522: GSM8174420: Arabinogalactan, KHP1, biol rep 3; Xylanibacter ruminicola; RNA-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 7.8M spots, 2.3G bases, 732.6Mb downloads

External Id: GSM8174420_r1
Submitted by: Biotechnical Faculty
Study: Transcriptomic analysis of polysaccharide utilization loci reveals substrate preferences in two ruminal generalists Segatella bryantii TF1-3 and Xylanibacter ruminicola KHP1
show Abstracthide Abstract
Bacteria of the genera Xylanibacter and Segatella are one of the most dominant groups in the rumen microbiota. They are characterized by the ability to utilize different hemicelluloses and pectin of plant cell-wall as well as plant energy storage polysaccharides. The degradation is possible with the use of cell envelope bound multiprotein apparatuses coded in polysaccharide utilization loci (PULs), which have been shown to be substrate specific. The knowledge of PUL presence in rumen Xylanibacter and Segatella based on bioinformatic analyses is already established and transcriptomic and genetic approaches confirmed predicted PULs for a limited number of substrates. In this study, we transcriptomically identified additional different PULs in Xylanibacter ruminicola KHP1 and Segatella bryantii TF1-3. We also identified substrate preferences and found that specific growth rate and extent of growth impacted the choice of substrates preferentially used for degradation. These preferred substrates were used by both strains simultaneously as judged by their PUL upregulation. Lastly, ß-glucan and xyloglucan were used by these strains in the absence of bioinformatically and transcriptomically identifiable PUL systems. Overall design: We were growing each bacteria in 11 different polysaccharides and glucose as reference. And performed RNAseq analysis for each polysaccharide (22 experiments) + 2 glucose. We also did the RNAseq in minimal medium with glucose and beta-glucan for S. bryantii TF1-3.
Sample: Arabinogalactan, KHP1, biol rep 3
SAMN40648913 • SRS20884801 • All experiments • All runs
Library:
Name: GSM8174420
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Strains were grown in liquid minimal or modified DSMZ medium 330 containing different substrates, until they reached OD value of 0.3-0.5. 1.5 mL of each triplicate was removed and immediately combined with 3 mL of RNAprotect bacteria reagent (Qiagen). RNA was purified using RNeasy Mini Kit (Qiagen) with the on-column DNase treatment. The quantity of the isolated RNA was determined using Spark 10M multifunctional microplate reader with Quant-iTTM RNA Assay Kit (Thermo Scientific). Illumina stranded TruSeq RNA library incl bacterial ribodepletion or NEBNext Ultra Directional RNA library prep kit
Runs: 1 run, 7.8M spots, 2.3G bases, 732.6Mb
Run# of Spots# of BasesSizePublished
SRR284933017,754,9652.3G732.6Mb2024-05-08

ID:
32403803

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