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在无核酸酶的T4多核苷酸激酶中出现的一种核酸内切酶活性的部分特性

Partial characterization of an endonuclease activity which appears in nuclease free T4 polynucleotide kinase.

作者信息

Loewen P C

出版信息

Nucleic Acids Res. 1976 Nov;3(11):3133-41. doi: 10.1093/nar/3.11.3133.

DOI:10.1093/nar/3.11.3133
PMID:188016
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC343157/
Abstract

A nuclease activity has been found to appear in preparations of T4 induced polynucleotide kinase which had originally been nuclease free. The nuclease introduced random nicks into T7 DNA suggesting that it was an endonuclease. Destabilization of the kinase molecule by osmotic shock or by the removal of reducing agents, ATP or salts was shown to stimulate the endonuclease appearance. The molecular weight was found to be 32,000 +/- 10% by gel filtration on G100 Sephadex. The nuclease was active over a wide pH range from pH 5.0 to pH 9.2 in a number of buffer systems and required MgCl2 and reducing agent for maximum activity. Sodium azide did not affect the nuclease appearance.

摘要

在最初不含核酸酶的T4诱导多核苷酸激酶制剂中发现出现了一种核酸酶活性。该核酸酶在T7 DNA上引入随机切口，表明它是一种内切核酸酶。通过渗透休克或去除还原剂、ATP或盐使激酶分子不稳定，结果显示会刺激内切核酸酶的出现。通过在G100 Sephadex上进行凝胶过滤，发现其分子量为32,000±10%。在多种缓冲系统中，该核酸酶在pH 5.0至pH 9.2的宽pH范围内具有活性，并且最大活性需要MgCl2和还原剂。叠氮化钠不影响核酸酶的出现。

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