Abstract:
:Several lines of evidence indicate that differences in DNA repair capacity are an important source of variability in cancer risk. However, traditional assays for measurement of DNA repair activity in human samples are laborious and time-consuming. DNA glycosylases are the first step in base excision repair of a variety of modified DNA bases. Here, we describe the development of a new sensitive DNA glycosylase assay based on fluorescence cross-correlation spectroscopy (FCCS) with two-photon excitation. FCCS was applied to the measurement of uracil DNA glycosylase activity of human cell extracts and validated by comparison with standard gel electrophoresis assay. Our results indicate that FCCS can be adapted to efficient assays for DNA glycosylase activity in protein extracts from human cells. This method has a potential for the development of automated screening of large number of samples.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Collini M,Caccia M,Chirico G,Barone F,Dogliotti E,Mazzei Fdoi
10.1093/nar/gni166keywords:
subject
Has Abstractpub_date
2005-10-21 00:00:00pages
e165issue
19eissn
0305-1048issn
1362-4962pii
33/19/e165journal_volume
33pub_type
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