In vitro detection of methylated DNA via recombinant protein MBD2b

Members of the methyl binding domain (MBD) protein family are known for binding to methylated DNA by recognizing methylated cytosines. Their original function is to regulate protein biosynthesis by recruitment of transcriptional repression complexes to silence gene expression. The aim of the present...

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Bibliographic Details
Published in:Molecular biology reports 2009-09, Vol.36 (7), p.1859-1862
Main Authors: Födermayr, Mathilde, Pröll, Johannes, Zach, Otto, Wechselberger, Christian, Lutz, Dieter
Format: Article
Language:English
Subjects:
Animal Anatomy
Animal Biochemistry
Antibodies - metabolism
Biomedical and Life Sciences
Deoxyribonucleic acid
DNA
DNA Methylation
DNA-Binding Proteins - isolation & purification
DNA-Binding Proteins - metabolism
Gene expression
Genomics
Histology
Life Sciences
Molecular biology
Morphology
Protein Binding
Proteins
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
Sequence Analysis, DNA - methods
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Description
Summary:Members of the methyl binding domain (MBD) protein family are known for binding to methylated DNA by recognizing methylated cytosines. Their original function is to regulate protein biosynthesis by recruitment of transcriptional repression complexes to silence gene expression. The aim of the presented work was to detect methylated DNA spotted onto nitrocellulose membranes with recombinant proteins MBD2b, MBD2b-GFP and directly labeled protein MBD2b. Proteins were affinity purified and tested for functionality before application. We were able to show that these functional recombinant proteins bind to unilaterally and symmetrically methylated oligonucleotides and genomic DNA in vitro and thus can be used in various detection assays.
ISSN:0301-4851
1573-4978
DOI:10.1007/s11033-008-9391-4