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
Jazyk: angličtina
Rok vydání: 2020
Předmět:
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