Ontology highlight
ABSTRACT:
PROVIDER: PRJNA633876 | ENA |
REPOSITORIES: ENA
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Project description:Whole-genome sequencing is an important way to understand the genetic information, gene function, biological characteristics, and living mechanisms of organisms. There is no difficulty to have mega-level genomes sequenced at present. However, we encountered a hard-to-sequence genome of Pseudomonas aeruginosa phage PaP1. The shotgun sequencing method failed to dissect this genome. After insisting for 10 years and going over 3 generations of sequencing techniques, we successfully dissected the PaP1 genome with 91,715 bp in length. Single-molecule sequencing revealed that this genome contains lots of modified bases, including 51 N6-methyladenines (m6A) and 152 N4-methylcytosines (m4C). At the same time, further investigations revealed a novel immune mechanism of bacteria, by which the host bacteria can recognize and repel the modified bases containing inserts in large scale, and this led to the failure of the shotgun method in PaP1 genome sequencing. Strategy of resolving this problem is use of non-library dependent sequencing techniques or use of the nfi- mutant of E. coli DH5M-NM-1 as the host bacteria to construct the shotgun library. In conclusion, we unlock the mystery of phage PaP1 genome hard to be sequenced, and discover a new mechanism of bacterial immunity in present study. Methylation profiling of Pseudomonas aeruginosa phage PaP1 using kinetic data generated by single-molecule, real-time (SMRT) sequencing on the PacBio RS.
2013-08-22 | E-GEOD-50100 | biostudies-arrayexpress
Project description:To detect the modifed bases in SINEUP RNA, we compared chemically modified in vitro transcribed (IVT) SINEUP-GFP RNA and in-cell transcribed (ICT) SINEUP RNA from SINEUP-GFP and sense EGFP co-transfected HEK293T/17 cells. Comparative study of Nanopore direct RNA sequencing data from non-modified and modified IVT samples against the data from ICT SINEUP RNA sample revealed modified k-mers positions in SINEUP RNA in the cell.
2023-02-04 | GSE224018 | GEO
Project description:SHAPE-MaP structure probing experiment was performed on SARS-CoV-2 infected Vero cells at 4 days post infection with two biological replicates. For each replciate, SHAPE-MaP includes a sample treated with 2-methylnicotinic acid imidazolide acid (modified) or a minue reagent (unmodified). NAI preferentially reacts with unpaired bases in RNA, forming acylated bases. These modifications are encoded as mutation during reverse transcripatse and library preparation. After sequencing and alignment, the reactivity profiles of 'modified' and 'unmodified' samples are used to calculate SHAPE reactivity of each base
2021-01-11 | GSE154171 | GEO
Project description:Here we report the analysis of the transcriptome of bone marrow-derived dendritic cells exposed to the different antigen delivery systems to dissect the molecular mechanisms that orchestrate the immune response upon vaccination. RNA-Sequencing revealed that exposure of dendritic cells to diffent antigen display vehicules causes the deregulation of molecules involved in the immune response, at a different extent, depending on the carrier. In vaccinology, patterns of gene expression induced by priming antigen presenting cells (APCs), could be used to predict immunogenicity and the type of T-helper cell polarization, and/or to helpin selecting an appropriate adjuvant to administer in the vaccine formulation. Bone marrow precursor at day 7 of GM-CSF culture were exposed or not to LPS deprived bacteriophage, bacteriophage modified to be targeted to DC or to the E2 protein scaffold
2015-03-01 | E-GEOD-60231 | biostudies-arrayexpress
Project description:Examination of modified bases on the genome of Ralstonia solanacearum UY031
2016-12-29 | GSE92982 | GEO
Project description:Partial exon deletion in Trpm7 gene was confirmed by lack of transcripts according to the RNA sequencing in two stem cell clones. For this mice were genetically modified by partial deletion of exon 2 and embryonic stem (ES) cell lines were generated. Isolated RNA of two ES clones was sequenced with Illumina NextSeq 500 Sequencer, which generated 75 bases long ‘single-end’ reads.
2021-09-06 | E-MTAB-10877 | biostudies-arrayexpress
Project description:Chemical probing has the power to provide insight into RNA conformation in vivo and in vitro, but interpreting the results depends on methods to detect the chemically modified nucleotides. Traditionally, the presence of modified bases was inferred from their ability to halt reverse transcriptase during primer extension and the locations of termination sites observed by electrophoresis or sequencing. More recently, modification-induced mutations have been used as a readout for chemical probing data. Given variable propensity for mismatch incorporation and read-through with different reverse transcriptases, we examined how termination and mutation events compare to each other in the same chemical probing experiments. We found that mutations and terminations induced by dimethyl sulfate probing are both specific for methylated bases, but these two measures have surprisingly little correlation and represent largely non-overlapping indicators of chemical modification data. We also show that specific biases for modified bases depend partly on local sequence context, and that different reverse transcriptases show different biases toward reading a modification as a stop or a mutation. These results support approaches that incorporate analysis of both termination and mutation events into RNA probing experiments.
2017-09-26 | GSE97609 | GEO
Project description:Identification of direct targets and modified bases of RNA cytosine methyltransferases
| PRJNA169525 | ENA
Project description:YerA41 is a myoviridae bacteriophage that was originally isolated due its ability to infect Yersinia ruckeri bacteria, the causative agent of enteric redmouth disease of salmonid fish. Several attempts to determine its genomic DNA sequence using traditional and next generation sequencing technologies failed, indicating that the phage genome is modified such way that it is an unsuitable template for PCR amplification and sequencing. To determine the YerA41 genome sequence we isolated RNA from phage-infected Y. ruckeri cells at different time points post-infection, and sequenced it. The host-genome specific reads were substracted and de novo assembly was performed on the unaligned reads.
2020-03-04 | GSE146319 | GEO
Project description:We use Illumina sequencing to monitor mutations in the bacteriophage T7 genome in the presence of T7 DNA polymerase that has an altered exonuclease active site. These alterations include mutation of key residues in the exonuclease active site.
2020-01-01 | GSE138098 | GEO