E. coli Topoisomerase IV Relaxation Assay Kits
These kits contain everything required for relaxation reactions E. coli Topoisomerase IV enzyme, the supercoiled pBR322 DNA substrate, the Assay and Dilution buffers as well as 10X GSTEB - our new stop and loading buffer.
DNA Topoisomerase IV (Topo IV) is a bacterial type II topoisomerase which can decatenate and relax supercoiled DNA and it is a validated target for antibacterial compounds, such as the quinolones and the coumarins. Escherichia coli Topo IV is prepared by overexpressing the ParC and ParE subunits in E. coli and purifying them by methods adapted from Peng and Marians, 19991.
1 U of topo IV will relax 0.5 µg supercoiled pBR322 DNA in 1X Assay Buffer in a 30 µL reaction volume in 30 minutes at 37°C.
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References
- Peng, H. and Marians, K.J. (1999) Overexpression and purification of bacterial topoisomerase IV, in DNA Topoisomerase Protocols Vol. I (Bjornsti, M-A., and Osheroff, N. eds.), Humana Press, Totowa, N.Jersey pp.163-169
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