By Osheroff N., Bjornsti M.A.

An unparalleled selection of state-of-the-art experimental protocols for investigating the catalytic actions of DNA topoisomerases, in addition to their particular interactions with topoisomerase-targeted antitumor and antibacterial medicines. defined by way of specialist experimentalists who've perfected the recommendations, those unfailingly reproducible equipment contain assays for enzyme-catalyzed DNA relaxation/supercoiling, DNA cleavage, DNA nicking, DNA decatenation, and ATP hydrolysis. numerous converted DNA substrates, used to dissect enzyme mechanisms through trapping intermediates, also are defined. Methodologies to figure out the motion of topoisomerase-targeted medications contain biochemical assays of drug-induced enzyme-DNA complexes, equipment for assaying drug uptake, and cell-based assays for identifying the specificity and mechanisms of drug resistance. A better half quantity, DNA Topoisomerase Protocols, I: DNA Topology and Enzymes, offers state of the art experimental protocols for investigating DNA constitution, topology, and DNA topoisomerase functionality.

Show description

Read or Download DNA Topoisomerase Protocols PDF

Similar genetics books

The Genetics of Renal Disease

Guy's medical institution, London, united kingdom. offers a finished account of the hereditary nephropathies and extra generalized problems that could impact the renal tract. formerly released because the Genetics of Renal Tract problems, by way of M. D'A Crawfurd, c1988. For scientific geneticists and researchers. Illustrated.

Molecular Genetics Medicine

Carrying on with to maintain speed with development in human molecular genetics, quantity four of Molecular Genetic drugs stories 5 new parts of serious significance. bankruptcy 1 reports the molecular mechanisms that experience beenunraveled within the pathogenesis of eye illnesses. the second one bankruptcy explains the extraordinary new precept if genomic imprinting, or epigenetic amendment imposed by means of parental background.

Extra info for DNA Topoisomerase Protocols

Example text

I. (1990) Archaebacterial reverse gyrase cleavage-site specificity is similar to that of eubacterial DNA topoisomerases I. Nucleic Acids Res. 18, 2801–2805. 21. Nadal, M. (1990) The reverse gyrase of Sulfolobus: DNA positive supercoiling in vitro and in vivo. D. thesis, Paris. PB Duguet et al. Topoisomerase II Hydrolysis 51 5 Topoisomerase II-Catalyzed ATP Hydrolysis as Monitored by Thin-Layer Chromatography Paul S. Kingma, John M. Fortune, and Neil Osheroff 1. Introduction Although type I topoisomerases do not require a high-energy cofactor, type II topoisomerases require ATP in order to carry out their essential catalytic functions (1–4).

Acta 1216, 213–220. 11. Barzilai, R. (1973) SV40 DNA: quantitative conversion of closed circular to open circular form by an ethidium bromide-restricted endonuclease. J. Mol. Biol. 74, 739–742. 12. Nakasu, S. and Kikuchi, A. (1985) Reverse gyrase; ATP-dependent type I topoisomerase from Sulfolobus. EMBO J. 4, 2705–2710. 13. , and Duguet, M. (1988) Reverse gyrase of Sulfolobus: purification to homogeneity and characterization. Biochemistry 27, 9102–9108. 14. Peck, L. J. and Wang, J. C. (1983) Energetics of B-to-Z transition in DNA.

Louis, MO, cat. #340-102). A vial with 2 mg contains enough NADH for 20 × 1 mL reactions. 4 mg of the PEP to a 2-mg vial of NADH. 0. This makes a 100× stock solution containing 16 mM NADH and 200 mM PEP (see Note 1). 3. Pyruvate Kinase and Lactate Dehydrogenase These two enzymes can be purchased as a mixture stored in 50% glycerol (Sigma #P-0294). A 5-mL bottle has enough enzyme for approx 750 × 1 mL reactions. 3. ATP Starting with the disodium salt, a stock solution of buffered ATP is made by dissolving 61 mg into 1 mL of double-distilled water.

Download PDF sample

Rated 4.93 of 5 – based on 16 votes