E. coli Gyrase

DNA gyrase is a bacterial type II topisomerase 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 from the over producing strains JMtacA and JMtacB¹ and is supplied as an A₂B₂ complex.

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
G1001	E.coli gyrase100 U	£110
G5002	E.coli gyrase500 U	£405
G1003	E.coli gyrase1000 U	£784
G2004	E.coli gyrase2000 U	£1,505

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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
K0001	E.coli Gyrase Supercoiling Kit 1100 U of E. coli gyrase and 50 µg of relaxed DNA	£191
K0002	E.coli Gyrase Supercoiling Kit 2500 U of E. Coli gyrase and 250 µg of relaxed DNA	£725
K003	E.coli Gyrase Supercoiling Kit 31000 U of E. coli gyrase and 500 µg of relaxed DNA	£1,447
K004	E.coli Gyrase Supercoiling Kit 42000 U of E. coli gyrase and 1 mg of relaxed DNA	£2,737

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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 enzyme and linearised pBR322 marker.

Cleavage specific enzyme available separately on request.

Technical Documents

Cat No.	Product	Price	Quantity
GCK001	E.coli Gyrase Cleavage Kit 1100 U of E. coli gyrase and 50 µg of supercoiled DNA	£376
GCK002	E.coli Gyrase Cleavage Kit 2500 U of E. coli gyrase and 250 µg of supercoiled DNA	£1,065
GCK003	E.coli Gyrase CleavageKit 31000 U of E. coli gyrase and 500 µg of supercoiled DNA	£1,470
GCK004	E.coli Gyrase CleavageKit 42000 U of E. coli gyrase and 1 mg of supercoiled DNA	£2,939

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These kits are designed for assaying cell extracts containing E. coli 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
GCE001	E.coli Cell Extract Assay Kit 1For 100 E. coli gyrase assays	£136
GCE002	E.coli Cell Extract Assay Kit 2For 500 E. coli gyrase assays	£402
GCE003	E.coli Cell Extract Assay Kit 3For 1000 E. coli gyrase assays	£799
GCE004	E.coli Cell Extract Assay Kit 4For 2000 E. coli gyrase assays	£1,527

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These kits can be used to test the effects of potential ATPase inhibitors. For example, the coumarin drugs such as novobiocin inhibit the action of DNA gyrase by competitively inhibiting the hydrolysis of ATP thus preventing supercoiling. 

These assays are microtitre plate-based and thus large numbers of compounds can be screened in a relatively short period of time. They also continuous assays which can provide more information than an end point assay.

Technical Documents

Cat No.	Product	Price	Quantity
ATPG001	Kit for 100 ATPase assaysFor determination of DNA-dependent ATPase activity; contains E. coli gyrase, buffers, ATP, PK-LDH, PEP, NADH and linear pBR322 DNA substrate.	£1,546
ATPG002	Kit for 100 ATPase assaysFor determination of DNA-independent ATPase activity; contains E. coli GyrB43 domain, buffers, PK-LDH, PEP, NADH and ATP.	£1,546

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The kit is supplied with sufficient E. coli gyrase enzyme, plasmid DNA substrate, buffers and other assay components for 100 assays. The enzyme is supplied at a concentration of 5 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
TRG01	HTS Assay Kit - E. coli Gyrase 1 - 5 KitsFor 100 assays per plate	£399
TRG02	HTS Assay Kit - E. coli Gyrase 6+ KitsFor 100 assays per plate	£340

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