Abstract:
:The role of uracil in genomic DNA has been recently re-evaluated. It is now widely accepted to be a physiologically important DNA element in diverse systems from specific phages to antibody maturation and Drosophila development. Further relevant investigations would largely benefit from a novel reliable and fast method to gain quantitative and qualitative information on uracil levels in DNA both in vitro and in situ, especially since current techniques does not allow in situ cellular detection. Here, starting from a catalytically inactive uracil-DNA glycosylase protein, we have designed several uracil sensor fusion proteins. The designed constructs can be applied as molecular recognition tools that can be detected with conventional antibodies in dot-blot applications and may also serve as in situ uracil-DNA sensors in cellular techniques. Our method is verified on numerous prokaryotic and eukaryotic cellular systems. The method is easy to use and can be applied in a high-throughput manner. It does not require expensive equipment or complex know-how, facilitating its easy implementation in any basic molecular biology laboratory. Elevated genomic uracil levels from cells of diverse genetic backgrounds and/or treated with different drugs can be demonstrated also in situ, within the cell.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Róna G,Scheer I,Nagy K,Pálinkás HL,Tihanyi G,Borsos M,Békési A,Vértessy BGdoi
10.1093/nar/gkv977subject
Has Abstractpub_date
2016-02-18 00:00:00pages
e28issue
3eissn
0305-1048issn
1362-4962pii
gkv977journal_volume
44pub_type
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