E. coli Gyrase

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E. coli Gyrase

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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 gyrase
100 U
£105
G5002 E.coli gyrase
500 U
£386
G1003 E.coli gyrase
1000 U
£746
G2004 E.coli gyrase
2000 U
£1,433
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E. coli Gyrase Supercoiling Assay Kits

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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 1
100 U of E. coli gyrase and 50 µg of relaxed DNA
£182
K0002 E.coli Gyrase Supercoiling Kit 2
500 U of E. Coli gyrase and 250 µg of relaxed DNA
£690
K003 E.coli Gyrase Supercoiling Kit 3
1000 U of E. coli gyrase and 500 µg of relaxed DNA
£1,378
K004 E.coli Gyrase Supercoiling Kit 4
2000 U of E. coli gyrase and 1 mg of relaxed DNA
£2,607
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E. coli Gyrase Cleavage Assay Kits

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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 1
100 U of E. coli gyrase and 50 µg of supercoiled DNA
£358
GCK002 E.coli Gyrase Cleavage Kit 2
500 U of E. coli gyrase and 250 µg of supercoiled DNA
£1,014
GCK003 E.coli Gyrase Cleavage Kit 3
1000 U of E. coli gyrase and 500 µg of supercoiled DNA
£1,400
GCK004 E.coli Gyrase Cleavage Kit 4
2000 U of E. coli gyrase and 1 mg of supercoiled DNA
£2,799
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E. coli Gyrase Assay Kits for Cell Extracts

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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 1
For 100 E. coli gyrase assays
£130
GCE002 E.coli Cell Extract Assay Kit 2
For 500 E. coli gyrase assays
£383
GCE003 E.coli Cell Extract Assay Kit 3
For 1000 E. coli gyrase assays
£761
GCE004 E.coli Cell Extract Assay Kit 4
For 2000 E. coli gyrase assays
£1,454
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E. coli Gyrase ATPase Kits

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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 assays
For determination of DNA-dependent ATPase activity; contains E. coli gyrase, buffers, ATP, PK-LDH, PEP, NADH and linear pBR322 DNA substrate.
£1,472
ATPG002 Kit for 100 ATPase assays
For determination of DNA-independent ATPase activity; contains E. coli GyrB43 domain, buffers, PK-LDH, PEP, NADH and ATP.
£1,472
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High / Medium-Throughput Assay Kit - E. coli Gyrase

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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 Kits
For 100 assays per plate
£390
TRG02 HTS Assay Kit - E. coli Gyrase 6+ Kits
For 100 assays per plate
£325
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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
  2. Maxwell, A., Burton, N.P. and O'Hagan, N. (2006) High-throughput assays for DNA gyrase and other topoisomerases. Nucleic Acid Res. 34 (15), e104