S. aureus Gyrase

Staphylococcus aureus gyrase is a bacterial type II topoisomerase which 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. It is prepared by overexpressing the subunits in E. coli and then purified.

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.

Technical Documents

Cat No.	Product	Price	Quantity
SAG4001	S. aureus gyrase100 U	£110
SAG4002	S. aureus gyrase500 U	£405

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Kits are available containing everything required to perform supercoiling reactions and to test inhibitors. They contain the enzyme, relaxed DNA substrate and the Assay and Dilution buffers.

Technical Documents

Cat No.	Product	Price	Quantity
SAS4001	S.aureus Gyrase Supercoiling Assay Kit 1100 U of S. aureus gyrase and 50 µg of relaxed DNA	£191
SAS4002	S.aureus Gyrase Supercoiling Assay Kit 2500 U of S. aureus gyrase and 250 µg of relaxed DNA	£704

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Some drugs interrupt the DNA breakage-reunion step of the gyrase reaction. This leads to cell death and it is the mechanism behind the action of the quinolones such as nalidixic acid and ciprofloxacin. Cleavage assays are particularly useful in determining if a potential drug acts by this mechanism.

These kits are designed specifically for cleavage reactions. They contain the supercoiled pBR322 DNA substrate and the Assay and Dilution buffers required for DNA cleavage reactions in addition to the S. aureus gyrase and linearised pBR322 marker.

Cleavage specific enzyme available separately on request.

Technical Documents

Cat No.	Product	Price	Quantity
SAGC001	S.aureus Gyrase Cleavage Assay Kit 1100 U of S. aureus gyrase and 50 ug of supercoiled DNA	£376
SAGC002	S.aureus Gyrase Cleavage Assay Kit 2500 U of S. aureus gyrase and 50 ug of supercoiled DNA	£1,065

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These kits are designed for assaying cell extracts containing gyrase and partially purified fractions and contain relaxed DNA substrate, Assay buffer, Dilution buffer and control supercoiled DNA. 

Technical Documents

Cat No.	Product	Price	Quantity
SAGE001	S.aureus Gyrase Cell Extract Assay Kit 1For 100 S. aureus gyrase assays	£136
SAGE002	S.aureus Gyrase Cell Extract Assay Kit 2For 500 S. aureus gyrase assays	£428

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The coumarin drugs inhibit the action of gyrase by competitively inhibiting the hydrolysis of ATP thus preventing supercoiling. These kits can be used to test the effects of potential ATPase inhibitors.

These kits are advantageous because the assays are microtitre plate-based and thus large numbers of compounds can be screened in a relatively short period of time. 

Technical Documents

Cat No.	Product	Price	Quantity
ATPSAG001	Kit for 100 ATPase assaysFor determination of DNA-dependent ATPase activity; contains S. aureus gyrase, buffers, ATP, PK-LDH, PEP, NADH and linear pBR322 DNA substrate.	£1,546

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The kit is supplied with sufficient S. aureus gyrase enzyme, plasmid DNA substrate, buffers and other assay components* for 100 assays. The enzyme is supplied at a concentration of 10 U/μl in Dilution Buffer. The kit is also supplied with sufficient wash buffers for one 96-well plate. These buffers are supplied as 20X concentrates and must be diluted with ultra pure water prior to use.

More information about this assay can be found on the "Services" page under "High/Medium Throughput Assay".

Kit issued with limited licence for individual use only. 

Patent held by Inspiralis Ltd., Norwich, Norfolk, UK. (Patent No. GB0424953.8, US7838230)

Technical Documents

Cat No.	Product	Price	Quantity
SATRG01	HTS Assay Kit S. aureus Gyrase 1 - 5 KitsFor 100 assays per plate	£410
SATRG02	HTS Assay Kit S. aureus Gyrase 6+ KitsFor 100 assays per plate	£340

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