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SRX9806232: GSM5009683: G. diazotrophicus cells, 0 µM FeCl3 R2; Gluconacetobacter diazotrophicus; RNA-Seq
1 ION_TORRENT (Ion Torrent PGM) run: 6.7M spots, 508.7M bases, 380.9Mb downloads

Submitted by: NCBI (GEO)
Study: Transcriptomic Response of the Diazotrophic Bacteria Gluconacetobacter diazotrophicus Strain PAL5 to Iron Limitation and Characterization of the fur Regulatory Network
show Abstracthide Abstract
Gluconacetobacter diazotrophicus has been the focus of several studies aiming to understand the mechanisms behind this endophytic diazotrophic bacterium. The present study is the first global analysis of the early transcriptional response of exponentially growing G. diazotrophicus to iron, an essential cofactor for many enzymes involved in various metabolic pathways. RNA-seq, targeted gene mutagenesis and computational motif discovery tools were used to define the G. diazotrophicus Fur regulon. The data analysis showed that genes encoding functions related to iron homeostasis, were significantly upregulated in response to iron limitation. Certain genes involved in the secondary metabolism were overexpressed under iron-limited conditions. In contrast, it was observed that the expression of genes involved in Fe-S cluster biosynthesis, flagellar biosynthesis and type IV secretion systems were downregulated in an iron-depleted culture medium. Our results support a model that control transcription in G. diazotrophicus by Fur function. The G. diazotrophicus Fur protein was able to complement an E. coli fur mutant. These results provide new insights into the effects of iron on the metabolism of G. diazotrophicus, as well as demonstrating the essentiality of this micronutrient for the main characteristic of plant growth promotion by G. diazotrophicus. Overall design: Profile mRNA obtained after half an hour incubation in the presence and absence of iron in triplicate.
Sample: G. diazotrophicus cells, 0 µM FeCl3 R2
SAMN17258057 • SRS7990222 • All experiments • All runs
Library:
Instrument: Ion Torrent PGM
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was isolated with Trizol in accordance with the manufacturer's protocol (Life Technologies) and treated with DNase I (Epicenter) in order to remove genomic DNA contamination. RNA purity was quantified using Qubit (Life Technologies). Seven micrograms of total RNA were used to ribosomal RNA (rRNA) depletion with MICROBExpressTM kit. The efficiency of depletion was evaluated in agarose gel electrophoresis (1%) followed by quantification of the total RNA with Agilent 2100 Bioanalyzer (Agilent). A total of 500 ng mRNA was used for the contruction of a sequencing library using the standard protocol of the SOLid Total RNAseq Kit (Life Technologies). The libraries were barcoded by using the SOLiD Transcriptome Multiplexing Kit (Life Technologies). The emulsion PCR and sequencing were performed according to Ion One TouchTM 200 Template Kit v2 DL and Ion PITM Sequencing 200 Kit v2 using the standard Life Technologies protocols, respectively. The Ion Proton Semiconductor Sequence (Life) was used to sequence twelve libraries generated from three biological replicates from each independent treatment.
Experiment attributes:
GEO Accession: GSM5009683
Links:
Runs: 1 run, 6.7M spots, 508.7M bases, 380.9Mb
Run# of Spots# of BasesSizePublished
SRR133869496,735,220508.7M380.9Mb2021-01-09

ID:
12840379

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