E. coli Gyrase Cleavage Assay Kits

These kits are designed specifically for DNA cleavage reactions. They contain cleavage specific enzyme, supercoiled pBR322 DNA substrate, the Assay and Dilution buffers, as well as 10X GSTEB - our new stop and loading buffer.

Interruption of the DNA breakage-reunion step or stabilisation of the DNA-breakage step of the gyrase reaction can lead to cell death and is the mechanism by which the quinolones, such as nalidixic acid and ciprofloxacin, act. Cleavage assays are particularly useful in determining if a potential compound acts by this mechanism. Quinolones and similar compounds do not require ATP to stabilise the cleavage intermediate, thus the cleavage assay buffer does not contain ATP. However, if you are concerned that your compound/molecule of interest requires ATP to stabilise the cleaved intermediate, ATP can be added to the reaction.

Cleavage specific enzyme is available separately on request.

DNA gyrase (gyrase) is a bacterial type II topoisomerase that can introduce negative supercoils into DNA. It is a target for both quinolone and coumarin drugs and can be used for screening potential antibacterial compounds. Escherichia coli DNA gyrase is prepared from the over producing strains JMtacA and JMtacB¹ and is supplied as an A₂B₂ complex.

Store at -80°C.

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References

1. Hallett, P., Grimshaw, A.J., Wigley, D.B. and Maxwell, A. (1990) Cloning of the DNA gyrase genes under tac promoter control: overproduction of the gyrase A and B proteins. Gene 93: 139-142