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Mycobacterium tuberculosis gyrase is a bacterial type II topoisomerase which can introduce negative supercoils into DNA. It is a target antibacterial drugs and can be used for screening potential novel antibacterial compounds. It is prepared by overexpressing the subunits with C-terminal polyhistidine tags in E. coli and then purified by affinity chromatography.
It is supplied as an A2B2 complex in Dilution Buffer. Store at -80 °C.
All enzyme is supplied with 5X concentrated Assay Buffer and Dilution Buffer which are also available separately. 1 U of gyrase will supercoil 0.5 µg relaxed pBR322 DNA in 30 minutes at 37 °C.
See technical documents below for more detailed information and lot specific activities.
Kits are available containing everything required to perform supercoiling reactions and to test inhibitors. They contain M. tuberculosis gyrase, relaxed DNA substrate and the Assay and Dilution buffers.
These contain the M. tuberculosis gyrase and the kDNA substrate in addition to the Assay and Dilution buffers for decatenation reactions. 1 U of M. tuberculosis gyrase will decatenate 200 ng of kDNA when incubated in 1X Assay buffer in a total reaction volume of 30 µl at 37°C for 30 minutes.
These kits are designed for assaying cell extracts containing M. tuberculosis gyrase and partially purified fractions and contain relaxed DNA substrate, Assay buffer, Dilution buffer and control supercoiled DNA.