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SRX10957118: GSM5329202: fnr21%-3; Komagataeibacter xylinus; RNA-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 8M spots, 2.4G bases, 706.8Mb downloads

Submitted by: NCBI (GEO)
Study: Effects of FNR, Arca and oxygen on the synthesis of bacterial cellulose
show Abstracthide Abstract
Purpose:The oxygen-regulated genes FNR and ARCA were combined with Komagataeibacter xylinus CGMCC 2955 to provide a new perspective for the study of the mechanism of oxygen environment on BC synthesis. Methods:The FNR and Arca overexpressing strains and the control strains were fermented under different partial oxygen pressures. The bacterial cellulose membrane in the logarithmic period of fermentation was enzymolyzed, and the bacteria were collected for transcriptome analysis.Sequencing was performed with Illumina and transcriptome analysis was performed on the bacteria under different conditions. Results:Transcriptome sequencing was performed using Illumina high-throughput sequencing technology on K. xylinus cultured under different oxygen tensions. The differentially expressed genes in the arcA overexpressing strains were mainly in the sulfur metabolism, two-component system, purine metabolism, and amino acid metabolism pathways compared to the control strains. Analysis showed that the arcA overexpression strain activated the sulfur metabolic pathway in K. xylinus. Due to the insufficient oxygen electron acceptors in the hypoxia, sulfate acted as the final electron acceptor and enhanced the growth ability of the strain. Through global regulation of the pathways of bacterial growth and metabolism as well as BC synthesis under low oxygen conditions, the arcA gene has enabled the strain to reach new levels of BC production. This study lays the foundation for further investigation of the mechanism of the effect of oxygen on BC synthesis in K. xylinus. Overall design: K. xylinus/ pseva331-FNR, K. xylinus/ pseva331-Arca and K. xylinus/pSEVA331 were cultured in static culture at 15%, 21% and 40% oxygen partial pressures and sequenced by Illumina high-throughput sequencing technology
Sample: fnr21%-3
SAMN19299741 • SRS9035388 • All experiments • All runs
Library:
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA was used as input material for the RNA sample preparations. For prokaryotic samples, mRNA was purified from total RNA using probes to remove rRNA. Fragmentation was carried out using divalent cations under elevated temperature in First Strand Synthesis Reaction Buffer(5X). First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase, then use RNaseH to degrade the RNA. And in the DNA polymerase I system, use dUTP to replace the dNTP of dTTP as the raw material to synthesize the second strand of cDNA. Remaining overhangs were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3' ends of DNA fragments, Adaptor with hairpin loop structure were ligated to prepare for hybridization. Then USER Enzyme was used to degrade the second strand of cDNA containing U, In order to select cDNA fragments of preferentially 370~420 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA). Then PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia) according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced on an Illumina Novaseq platform and 150 bp paired-end reads were generated.
Experiment attributes:
GEO Accession: GSM5329202
Links:
Runs: 1 run, 8M spots, 2.4G bases, 706.8Mb
Run# of Spots# of BasesSizePublished
SRR146150887,976,6792.4G706.8Mb2021-05-24

ID:
14574422

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