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首页> 外文期刊>Journal of bacteriology >Purification and partial characterization of the principal deoxyribonucleic acid polymerase from Mycoplasmatales.
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Purification and partial characterization of the principal deoxyribonucleic acid polymerase from Mycoplasmatales.

机译：来自支原体的主要脱氧核糖核酸聚合酶的纯化和部分表征。

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In this report we present the first description of the isolation and partial characterization of the deoxyribonucleic acid (DNA) polymerase activity from two species of Mycoplasmatales, Mycoplasma orale type 1 and M. hyorhinis. We have identified only a single DNA polymerase species in the mycoplasma crude extracts, and the enzymes from the two organisms are very similar in their structural and enzymatic properties. The purified polymerase from each source has a specific activity of greater than 50,000 U/mg of protein, a sedimentation coefficient of 5.6s, and an estimated molecular weight by gel filtration of 130,000. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the most highly purified M. orale fraction contains a single major protein band of 130,000 daltons, which we believe may represent the polymerase protein. The enzymes are most reactive with gapped (activated) DNA and show a marked preference for this primer template over oligodeoxyribonucleotide-initiated homoribo- or homodeoxyribo-polymers. The most purified preparations are devoid of contaminating endonuclease activity and also appear to lack associated 5' leads to 3'- or 3' leads to 5'-exonuclease activities, as determined by highly sensitive assays. The absence of the 3' leads to 5'-exonuclease is particularly remarkable in that this activity is essentially ubiquitous among the DNA polymerases that have thus far been characterized from procaryotes.
机译：在本报告中，我们首次介绍了从两种支原体种类（口头支原体和透明质酸支原体）中脱氧核糖核酸（DNA）聚合酶活性的分离和部分表征。我们在支原体粗提物中仅鉴定了一种DNA聚合酶，而这两种生物的酶在结构和酶学性质上都非常相似。来自每种来源的纯化的聚合酶的比活度均大于50,000 U / mg蛋白质，沉降系数为5.6s，通过凝胶过滤估算的分子量为130,000。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上，纯化度最高的M. orale馏分包含一个主蛋白质带，其分子量为130000道尔顿，我们认为这可能代表了聚合酶蛋白质。该酶与有缺口的（活化的）DNA最具反应性，并且比寡聚脱氧核糖核苷酸引发的同核糖或同脱氧核糖聚合物对这种引物模板表现出明显的偏好。最纯化的制剂没有污染核酸内切酶的活性，而且似乎缺乏相关的5'导致3'-或3'导致5'-核酸外切酶的活性，这是通过高灵敏度测定确定的。不存在3'导致5'-核酸外切酶是特别显着的，因为该活性在迄今已从原核生物中表征的DNA聚合酶中基本上是普遍存在的。

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