A novel decatenation assay for DNA topoisomerases using a singly-linked catenated substrate
Autor: | William Marshall Stark, Anthony Maxwell, Shruti Jain, Sean D. Colloms, Nidda F. Waraich, Nicolas P. Burton |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
DNA
Catenated DNA gyrase General Biochemistry Genetics and Molecular Biology Substrate Specificity 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine In vivo Enzyme Assays 030304 developmental biology Recombination Genetic 0303 health sciences Base Sequence biology Chemistry Topoisomerase DNA replication Substrate (chemistry) In vitro Biochemistry 030220 oncology & carcinogenesis Kinetoplast biology.protein Transposon Resolvases DNA Topoisomerases DNA Biotechnology |
ISSN: | 0736-6205 |
Popis: | Decatenation is a crucial in vivo reaction of DNA topoisomerases in DNA replication and is frequently used in in vitro drug screening. Usually this reaction is monitored using kinetoplast DNA as a substrate, although this assay has several limitations. Here we have engineered a substrate for Tn 3 resolvase that generates a singly-linked catenane that can readily be purified from the DNA substrate after restriction enzyme digestion and centrifugation. We show that this catenated substrate can be used with high sensitivity in topoisomerase assays and drug-inhibition assays. |
Databáze: | OpenAIRE |
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