Monkeypox Virus Topoisomerase 1 Enzyme

Monkeypox Virus topoisomerase 1 enzyme is supplied in Dilution Buffer1 at a minimum concentration of 30ng/µl, corresponding to at least 30 U/µl. 1 U of mvpTOP1 relaxes 0.5 µg of supercoiled pUC19 when incubated in 1xReaction Buffer2 and 150 mM NaCl for 30 minutes at 37°C – see Documentation.

The enzyme preparation is highly stable and recommended for activity assays, including cleavage-ligation experiments, equilibrium, nicking, and relaxation assays.

The enzyme is supplied together with:
• Dilution Buffer: 5 mM Tris-HCl pH 7.5, 400 mM NaCl, 1mM DTT, 50% glycerol
• Reaction Buffer 10x: 100 mM Tris-HCl pH 7.5; 50 mM MgCl2, 50 mM CaCl2; 50 µg/ml BSA
• Nucleases-free water
• 1M NaCl

Recombinant 6HIS-tagged monkeypox virus topoisomerase 1 is prepared by overexpressing the enzyme in Saccharomyces cerevisiae followed by purification by affinity chromatography adapted from Knudsen et al, J. Chrom. 1996, DOI: 10.1016/0378-4347(96)00152-1

The enzyme is shipped packed with ice gel packs and must be stored at -18°C to -20°C upon arrival. The temperature must be controlled to avoid that the enzyme preparation solidifies. This would destroy the enzyme integrity and cause a drop of the monkeypox virus topoisomerase 1 activity. The enzyme has a lifetime of 12 months when stored under optimal conditions

Certificate of Analysis for Monkeypox Virus topoisomerase 1

Safety Data Sheet for Monkeypox Virus topoisomerase 1 enzyme products

1. Estep, R.D.; Messaoudi, I.; O’Connor, M.A.; Li, H.; Sprague, J.; Barron, A.; Engelmann, F.; Yen, B.; Powers, M.F.; Jones, J.M.; et al. Deletion of the Monkeypox Virus Inhibitor of Complement Enzymes Locus Impacts the Adaptive Immune Response to Monkeypox Virus in a Nonhuman Primate Model of Infection. J. Virol. 2011, 85, 9527–9542, doi:10.1128/jvi.00199-11.

2. Shuman, S. Vaccinia virus DNA topoisomerase: A model eukaryotic type IB enzyme. Biochim. Biophys. Acta – Gene Struct. Expr. 1998, 1400, 321–337, doi:10.1016/S0167-4781(98)00144-4.

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