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ERX6822633: Illumina MiSeq paired end sequencing; Response of Leptospirillum ferriphilum DSM14647 to elevated NaCl concentration
1 ILLUMINA (Illumina MiSeq) run: 3.3M spots, 496.9M bases, 266.9Mb downloads

Design: Response of Leptospirillum ferriphilum DSM14647 to elevated NaCl concentration
Submitted by: Technische Universitat Bergakademie Freiberg Institute of Biosciences (Technische Universitat Bergakademie Freiberg Insti)
Study: Response of Leptospirillum ferriphilum DSM14647 to elevated NaCl concentration
show Abstracthide Abstract
Leptospirillum ferriphilum is an important acidophilic ferrous iron-oxidizing species for bioleaching or biooxidation to win metals like copper or gold, respectively. L. ferriphilum is inhibited by elevated conentrations of chloride. In the present study, the type strain of L. ferriphilum (i.e. strain DSM 14647) by subcultivation in presence of increasing chloride concentrations was adapted to tolerate higher concentrations of NaCl. The adapted culture was grown in presence of 180 mM NaCl or and the non-adapted culture without NaCl, and both were harvested in the late exponential phase. Total RNA was isolated and checked for quality and integrity. For each of the two conditions three RNA preparations of high quality (integrity number above 7) were pooled. Ribosomal RNA was depleted. DNA libraries for paired-end sequencing on an Illumina MiSeq were generated with a TruSeq stranded mRNA library prep kit (Illumina). Among the genes up-regulated were those coding for proteins likely involved in intracellular pH regulation, response to reactive oxygen species, and Fe-S cluster biosynthesis. Among the genes down-regulated in presence of chloride were those related to lipopolysaccharide and peptidoglykane synthesis, and interestingly also those for (hydroxy) ectoine biosynthesis.
Sample: Sample 2
SAMEA10838349 • ERS8489583 • All experiments • All runs
Library:
Name: Sample 2_p
Instrument: Illumina MiSeq
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: Inverse rRNA
Layout: PAIRED
Construction protocol: Centrifugation of culture in late exponential phase at 8000 x g and 4C for 15 minutes. Washing of cells with cold 10 mM H2SO4, and twice with 10 mM sodium citrate pH 7.0. Growth of L.ferriphulum in DSMZ 882 medium (pH1.8) supplemented with 20 g L-1 ferrous sulfate in a shaker at 37C and 180 rpm. Isolation of total RNA by RNeasy Mini Kit from Qiagen. DNA removed by treatment with DNase I (New England Biolabs). Evaluation of purity and integrity of total RNA with Agilent Bioanalyzer 2100 and an RNA 600 Nano Kit (Agilent Technologies. For each sample 3 RNA preparations with RMNA integrity number above 7 from independent biological replicates pooled into one. Depletion of ribosomal RNA by MICROBExpress kit (Thermo Fisher). Preparation of cDNA library by TruSeq stranded mRNA library prep kit (Illumina).
Experiment attributes:
Experimental Factor: compound: sodium chloride
Experimental Factor: dose: 180
Runs: 1 run, 3.3M spots, 496.9M bases, 266.9Mb
Run# of Spots# of BasesSizePublished
ERR72531723,310,082496.9M266.9Mb2022-04-02

ID:
21018916

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