E. coli Gyrase
- E. coli Gyrase
- E. coli Gyrase Supercoiling Assay Kits
- E. coli Gyrase Cleavage Assay Kits
- E. coli Gyrase Assay Kits for Cell Extracts
- E. coli Gyrase ATPase Kits
- High / Medium-Throughput Assay Kit - E. coli Gyrase
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.
The enzyme is supplied at a concentration of 5 U/µl 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.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| G1001 | E.coli gyrase | 100 U | £90 |
| G5002 | E.coli gyrase | 500 U | £330 |
| G1003 | E. coli gyrase | 1000 U | £645 |
| G2004 | E. coli gyrase | 2000 U | £1240 |
E. coli Gyrase Supercoiling Assay Kits
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.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| K0001 | Kit 1 | 100 U of E. coli gyrase and 50 µg of relaxed DNA | £155 |
| K0002 | Kit 2 | 500 U of E. coli gyrase and 250 µg of relaxed DNA | £595 |
| K0003 | Kit 3 | 1000 U of E. coli gyrase and 500 µg of relaxed DNA | £1190 |
| K0004 | Kit 4 | 2000 U of E. coli gyrase and 1 mg of relaxed DNA | £2250 |
E. coli Gyrase Cleavage Assay Kits
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.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| GCK001 | Kit 1 | 100 U of E. coli gyrase and 50 µg of supercoiled DNA | £310 |
| GCK002 | Kit 2 | 500 U of E. coli gyrase and 250 µg of supercoiled DNA | £875 |
| GCK003 | Kit 3 | 1000 U of E. coli gyrase and 500 µg of supercoiled DNA | £1210 |
| GCK004 | Kit 4 | 2000 U of E. coli gyrase and 1 mg of supercoiled DNA | £POA |
E. coli Gyrase Assay Kits for Cell Extracts
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.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| GCE001 | Kit 1 | for 100 E. coli gyrase assays | £110 |
| GCE002 | Kit 2 | for 500 E. coli gyrase assays | £350 |
| GCE003 | Kit 3 | for 1000 E. coli gyrase assays | £690 |
| GCE004 | Kit 4 | for 2000 E. coli gyrase assays | £1320 |
E. coli Gyrase ATPase Kits
The coumarin drugs inhibit the action of DNA 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.
| Cat No. | Product | Description | Price |
|---|---|---|---|
| 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. | £1270 |
| ATPB001 | Kit for 100 ATPase assays | For determination of DNA-independent ATPase activity; contains E. coli GyrB43 domain, buffers, PK-LDH, PEP, NADH and ATP. | £1270 |
High / Medium-Throughput Assay Kit - E. coli Gyrase
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.
* Please note: The recommended fluorescence dye for this kit is SYBR Gold ® (Invitrogen) NOT SUPPLIED
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 Plant Biosciences Ltd., Norwich, Norfolk, UK. ( Patent No. GB0424953.8)
| Cat No. | Product | Description | Price |
|---|---|---|---|
| TRG01 | HTS Assay Kit - E. coli Gyrase 1 - 5 Kits | for 100 assays per plate | £330 |
| TRG02 | HTS Assay Kit - E. coli Gyrase 6+ Kits | for 100 assays per plate | £260 |
References
- 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
- 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