Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1994-12-30
1997-08-12
Naff, David M.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
435233, 536 221, C12P 1934, C07H 2104, C12N 990
Patent
active
056564635
ABSTRACT:
A type 1 topoisomerase, designated topoisomerase V, has been isolated and substantially purified from the halophilic thermophilic methanogen bacterium Methanopyrus kandleri. The topoisomerase was purified by a process including the steps of lysing cells of M. kandleri to form a lysate, treating the lysate with polyethyleneimine to form a precipitate and a supernatant, precipitating the polyethyleneimine supernatant with ammonium sulfate, chromatographing the ammonium sulfate precipitate on phosphocellulose to produce a phosphocellulose eluate, chromatographing the phosphocellulose eluate on heparin to produce a heparin eluate, and chromatographing the heparin eluate on a column capable separating proteins by molecular size therein to produce a substantially purified thermostable DNA topoisomerase V. Topoisomerase V can relax DNA and can unlink DNA by reducing the linking number of closed circular DNA. Topoisomerase V is active over a wide range of temperatures and ionic conditions and does not require magnesium or ATP for its activity. It is recognized by anti-human topoisomerase I antibody.
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Fidelity Systems, Inc.
Naff David M.
Weber Jon P.
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