E. coli Topoisomerase IV

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 total reaction volume of 30 µl in 30 minutes at 37°C.

1 U of topo IV will decatenate 200 ng of kDNA when incubated in 1X Assay buffer in a total reaction volume of 30 µl in 30 minutes at 37°C. 

The ParC and ParE subunits are purified to >95% purity as judged by SDS-PAGE. The topo IV is supplied as a heterotetramer complex in Dilution buffer.

Store at -80ºC.

All enzyme is supplied with 5X concentrated Assay Buffer and Dilution buffers which are also available separately.

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References

1. 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