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[嗜热栖热菌YT-1中DNA聚合酶的分离及性质]

[Isolation and properties of DNA polymerase from extreme thermophylic bacteria Thermus aquaticus YT-1].

作者信息

Kaledin A S, Sliusarenko A G, Gorodetskiĭ S I

出版信息

Biokhimiia. 1980 Apr;45(4):644-51.

PMID:7378495
Abstract

A DNA polymerase has been isolated from the thermophylic bacteria Thermus aquaticus YT-1. The six-step purification procedure resulted in an electrophoretically homogeneous enzyme preparation with molecular weight of about 62 000. The enzyme does not contain contaminant exo- and endonuclease activities and has a temperature optimum on the DNA templates at 70 degress and that on RNA matrices at 50 degrees. The maximal activity of the enzyme requires the presence of bivalent cations (magnesium or manganese) 0,1--0,2 M KCl or NaCl, all deoxynucleoside triphosphates and template in the incubation mixture. The enzyme is active when "activated" DNA, poly(dA)-poly(dT), poly(dA)-oligo(dT)10 and poly(rA)-oligo(dT)10 are used as templates and is inactive on the native and denaturated DNAs as well as on the native molecules of RNA and poly(rC)-oligo(dG)12--180.

摘要

已从嗜热细菌水生栖热菌YT-1中分离出一种DNA聚合酶。经过六步纯化程序,得到了一种电泳纯的酶制剂,其分子量约为62000。该酶不含有污染性的外切核酸酶和内切核酸酶活性,在DNA模板上的最适温度为70℃,在RNA模板上为50℃。该酶的最大活性需要在孵育混合物中存在二价阳离子(镁或锰)、0.1 - 0.2M KCl或NaCl、所有脱氧核苷三磷酸和模板。当使用“活化”的DNA、聚(dA)-聚(dT)、聚(dA)-寡聚(dT)10和聚(rA)-寡聚(dT)10作为模板时,该酶具有活性,而对天然和变性的DNA以及天然RNA分子和聚(rC)-寡聚(dG)12 - 180无活性。

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