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SRX6451468: GSM3942645: OL161-ind-2; Hyphomonas neptunium; RNA-Seq
1 ILLUMINA (NextSeq 500) run: 15.4M spots, 1.2G bases, 399.7Mb downloads

Submitted by: NCBI (GEO)
Study: Analysis of global transcriptional changes in H. neptunium cells depleted of the cell cycle regulators CckA or ChpT
show Abstracthide Abstract
Within the bacterial phylum of the Alphaproteobacteria many species show complex life cycles. Proper regulation of these life cycles requires cell cycle regulation pathways, one of which is the so-called CtrA pathway. Key factors in the CtrA pathway are the histidine kinase CckA and the phosphotransferase ChpT, which are directly involved in cell cycle specific activation of the response regulator CtrA. Within the Alphaproteobacteria, stalked budding bacteria are even more asymmetric than most other representatives. How they regulate their cell cycle has not been studied before. Here, we investigated the transcriptional profiles of Hyphomonas neptunium cells depleted of CckA or ChpT and compared their profiles to that of wildtype cells. We identified that the essential proteins CckA and ChpT influence the transcription levels of multiple essential processes, such as cell division, as well as specific cell cycle regulated functions such as motility. Overall design: mRNA profiles for H. neptunium wild-type cells or cells depleted of CckA or ChpT were generated in duplicate by next-generation sequencing, using an Illumina NextSeq 500 system with 75 bp read length and a "High 75" sequencing kit
Sample: OL161-ind-2
SAMN12284354 • SRS5104873 • All experiments • All runs
Library:
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was isolated from the cell pellets using a bead mill and the mirVana RNA isolation kit (Ambion) including DNase treatment. From the total RNA samples, ribosomal RNA molecules were depleted using the Ribo-Zero rRNA Removal Kit for bacteria (Epicentre). From the rRNA-depleted RNA samples, first-strand cDNA was synthesized using a N6 randomized primer. After fragmentation, the Illumina TruSeq sequencing adapters (containing barcode sequences) were ligated in a strand-specific manner to the 5' and 3' ends of the cDNA fragments. The cDNA was finally amplified with PCR using a proof-reading enzyme. For Illumina NextSeq sequencing, the cDNAs were pooled in approximately equimolar amounts. The cDNA pool was eluted from a preparative agarose gel in the size range of 200 – 500 bp. The primers used for PCR amplification were designed for TruSeq sequencing according to the instructions of Illumina, yielding a total length of the flanking sequences of 122 bp.
Links:
Runs: 1 run, 15.4M spots, 1.2G bases, 399.7Mb
Run# of Spots# of BasesSizePublished
SRR969319415,403,3821.2G399.7Mb2022-01-02

ID:
8547769

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