A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.

2.50
Hdl Handle:
http://hdl.handle.net/10143/142370
Title:
A novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.
Authors:
Robertson, Adam B; Dahl, John A; Vågbø, Cathrine B; Tripathi, Pankaj; Krokan, Hans E; Klungland, Arne
Citation:
Nucleic acids research 2011, 39 (8):e55

Full metadata record

DC FieldValue Language
dc.contributor.authorRobertson, Adam Ben
dc.contributor.authorDahl, John Aen
dc.contributor.authorVågbø, Cathrine Ben
dc.contributor.authorTripathi, Pankajen
dc.contributor.authorKrokan, Hans Een
dc.contributor.authorKlungland, Arneen
dc.date.accessioned2011-09-15T08:01:29Z-
dc.date.available2011-09-15T08:01:29Z-
dc.date.issued2011-04-
dc.identifier.citationNucleic acids research 2011, 39 (8):e55en
dc.identifier.issn1362-4962-
dc.identifier.pmid21300643-
dc.identifier.doi10.1093/nar/gkr051-
dc.identifier.urihttp://hdl.handle.net/10143/142370-
dc.description.abstractRecently, 5-hydroxymethylcytosine (5hmC) was identified in mammalian genomic DNA. The biological role of this modification remains unclear; however, identifying the genomic location of this modified base will assist in elucidating its function. We describe a method for the rapid and inexpensive identification of genomic regions containing 5hmC. This method involves the selective glucosylation of 5hmC residues by the β-glucosyltransferase from T4 bacteriophage creating β-glucosyl-5-hydroxymethylcytosine (β-glu-5hmC). The β-glu-5hmC modification provides a target that can be efficiently and selectively pulled down by J-binding protein 1 coupled to magnetic beads. DNA that is precipitated is suitable for analysis by quantitative PCR, microarray or sequencing. Furthermore, we demonstrate that the J-binding protein 1 pull down assay identifies 5hmC at the promoters of developmentally regulated genes in human embryonic stem cells. The method described here will allow for a greater understanding of the temporal and spatial effects that 5hmC may have on epigenetic regulation at the single gene level.en
dc.language.isoenen
dc.subjectVDP::Medisinske Fag: 700::Basale medisinske, odontologiske og veterinærmedisinske fag: 710::Medisinsk genetikk: 714en
dc.subject.meshCarrier Proteinsen
dc.subject.meshCytosineen
dc.subject.meshDNAen
dc.subject.meshEmbryonic Stem Cellsen
dc.subject.meshGene Expression Regulationen
dc.subject.meshGenome, Humanen
dc.subject.meshGenomicsen
dc.subject.meshGlucosyltransferasesen
dc.subject.meshHumansen
dc.subject.meshPromoter Regions, Geneticen
dc.subject.meshProtozoan Proteinsen
dc.titleA novel method for the efficient and selective identification of 5-hydroxymethylcytosine in genomic DNA.en
dc.typeJournal articleen
dc.typepeer revieweden
dc.contributor.departmentCentre for Molecular Biology and Neuroscience and Institute of Medical Microbiology, Oslo University Hospital, Rikshospitalet, Norway.en
dc.identifier.journalNucleic acids researchen
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