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  • 标题:Purification of a DNA supercoiling factor from the posterior silk gland of Bombyx mori.
  • 本地全文:下载
  • 作者:T Ohta ; S Hirose
  • 期刊名称:Proceedings of the National Academy of Sciences
  • 印刷版ISSN:0027-8424
  • 电子版ISSN:1091-6490
  • 出版年度:1990
  • 卷号:87
  • 期号:14
  • 页码:5307-5311
  • DOI:10.1073/pnas.87.14.5307
  • 语种:English
  • 出版社:The National Academy of Sciences of the United States of America
  • 摘要:A protein factor with an estimated molecular mass of 50 kDa has been purified to homogeneity from the silk gland of Bombyx mori. In the presence of a molar excess of this factor and eukaryotic DNA topoisomerase II, relaxed circular DNA is converted to the negatively supercoiled form. Eukaryotic DNA topoisomerase I cannot substitute for eukaryotic DNA topoisomerase II in the supercoiling reaction. The reaction is dependent on ATP and is inhibited by VP-16, a specific inhibitor of eukaryotic DNA topoisomerase II. When DNA topoisomerase I is subsequently added to the supercoiling reaction mixture, the supercoiled DNA becomes relaxed. These results suggest that when both the 50-kDa protein and eukaryotic DNA topoisomerase II are present in excess, unconstrained negative supercoils are introduced into DNA.
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