Purification and characterization of Thermus thermophilus UvrD.
作者： Ruairi Collins , Tommie V. McCarthy
DOI: 10.1007/S00792-002-0293-4
关键词:
摘要: The DNA helicase UvrD (helicase II) protein plays an important role in nucleotide excision repair, mismatch rolling circular plasmid replication, and replication. A homologue of the Escherichia coli uvrD gene was previously identified Thermus thermophilus; however, to date, a has not been purified characterized from thermophile. Here we report purification characterization thermophilus HB8. temperature range 10° >65°C, with optimum 50°C, within limits assay. enzyme had requirement for divalent metal ions nucleoside triphosphates which related activity order ATP > dATP dGTP GTP >> CTP dCTP UTP. simple real-time assay developed that should facilitate detailed kinetic studies enzyme. Evaluation substrates using this showed highly active on double-stranded 5′ recessed ends comparison 3′ or blunt ends, supports translocation 3′–5′ direction relative strand bound by
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