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Merck
CN

T7428

Topoisomerase I from wheat germ

buffered aqueous glycerol solution

别名:

DNA relaxing enzyme, Topo I

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化学文摘社编号:
UNSPSC Code:
12352204
MDL number:
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grade

Molecular Biology

form

buffered aqueous glycerol solution

mol wt

97 kDa

foreign activity

RNase and DNase, none detected

shipped in

dry ice

storage temp.

−70°C

General description

Topoisomerase I relaxes supercoiled DNA molecules. The enzyme initiates transient breakages and rejoins of phosphodiester bonds in superhelical turns of closedcircular dsDNA.

Application

Suitable for:
  • Analsis of degree of DNA supercoiling
  • Forming knots and circles in single stranded DNA
  • Converting complementary single stranded DNA to double stranded, circular forms
  • Rejoining nicks in double-stranded DNA

Biochem/physiol Actions

Topoisomerase I relaxes supercoiled DNA molecules. The enzyme initiates transient breakages and rejoins of phosphodiester bonds in superhelical turns of closed-circular DNA. Enzyme activity is independent of right- and left-handed superhelices.

Other Notes

One unit will convert 1μg of supercoiled pGEM9Zf(−) plasmid DNA to the relaxed form in 30 min at 37 °C.
To preserve enzyme activity, avoid multiple freeze-thaw cycles and frequent temperature variations.
Topoisomerase I is supplied in a solution of 50 mM Tris-HCl, pH 7.9, 1 mM EDTA, 1 mM DTT, 0.5 M NaCl and 50% glycerol (v/v).

存储类别

11 - Combustible Solids

wgk

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)

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分析证书(COA)

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J C Wang
Annual review of biochemistry, 65, 635-692 (1996-01-01)
The various problems of disentangling DNA strands or duplexes in a cell are all rooted in the double-helical structure of DNA. Three distinct subfamilies of enzymes, known as the DNA topoisomerases, have evolved to solve these problems. This review focuses
P D Foglesong et al.
BioTechniques, 13(3), 402-404 (1992-09-01)
Supercoiled and relaxed DNA were resolved electrophoretically in the presence of 0.5 micrograms/ml ethidium bromide. Under these conditions the Gaussian distributions of topological isomers of both supercoiled and relaxed DNA migrated as discrete bands. The separation of these DNAs was
Lyudmila Yakovleva et al.
Biochemistry, 52(5), 984-991 (2013-01-16)
Vaccinia DNA topoisomerase IB (TopIB) relaxes supercoils by forming and resealing a covalent DNA-(3'-phosphotyrosyl(274))-enzyme intermediate. Conserved active site side chains promote the attack of Tyr274 on the scissile phosphodiester via transition state stabilization and general acid catalysis. Two essential side
Maris A Cinelli et al.
Journal of medicinal chemistry, 55(24), 10844-10862 (2012-12-12)
Hydroxylated analogues of the anticancer topoisomerase I (Top1) inhibitors indotecan (LMP400) and indimitecan (LMP776) have been prepared because (1) a variety of potent Top1 poisons are known that contain strategically placed hydroxyl groups, which provides a clear rationale for incorporating
Tomoko Nanbu et al.
Molecular and cellular biology, 33(6), 1175-1187 (2013-01-09)
Protection of telomeres protein 1 (Pot1) binds to single-stranded telomere overhangs and protects chromosome ends. RecQ helicases regulate homologous recombination at multiple stages, including resection, strand displacement, and resolution. Fission yeast pot1 and RecQ helicase rqh1 double mutants are synthetically

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