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嗜放射球菌属的质粒以及嗜放射栖热菌质粒pUE1的克隆与限制性图谱分析

The plasmids of Deinococcus spp. and the cloning and restriction mapping of the D. radiophilus plasmid pUE1.

作者信息

Mackay M W, al-Bakri G H, Moseley B E

出版信息

Arch Microbiol. 1985 Feb;141(1):91-4. doi: 10.1007/BF00446746.

Abstract

Plasmids were found in strains representing all four species of the genus Deinococcus viz. D. radiodurans, D. radiopugnans, D. radiophilus and D. proteolyticus but were not found in the most intensively-investigated strain of the genus, D. radiodurans R1. Their sizes were calculated from electron micrographs. D. radiophilus yielded three size classes of plasmid while D. radiodurans Sark, D. proteolyticus and D. radiopugnans each yielded two. Attempts to cure D. radiophilus and D. radiodurans Sark of any of their plasmids, using a variety of methods, were unsuccessful. A 10.8 kbase pair (kb) plasmid from D. radiophilus, pUE1, was cloned into the PstI site of pAT153 and propagated in Escherichia coli HB101. The recombinant plasmid, pUE109 was subjected to single and double digestion with various restriction endonucleases and its restriction map constructed. The resistance of E. coli HB101 to ultraviolet radiation was not increased when pUE109 was introduced into it. Attempts to transform D. radiodurans with pUE109 failed to detect tetracycline-resistant transformants.

摘要

在代表嗜放射菌属所有四个物种的菌株中发现了质粒,即耐辐射嗜放射菌、放射抗性嗜放射菌、嗜放射嗜放射菌和蛋白水解嗜放射菌,但在该属研究最深入的菌株耐辐射嗜放射菌R1中未发现。它们的大小通过电子显微镜照片计算得出。嗜放射嗜放射菌产生了三种大小类别的质粒,而耐辐射嗜放射菌Sark、蛋白水解嗜放射菌和放射抗性嗜放射菌各产生了两种。使用多种方法试图去除嗜放射嗜放射菌和耐辐射嗜放射菌Sark的任何一种质粒均未成功。来自嗜放射嗜放射菌的一个10.8千碱基对(kb)的质粒pUE1被克隆到pAT153的PstI位点,并在大肠杆菌HB101中繁殖。重组质粒pUE109用各种限制性内切酶进行单酶切和双酶切,并构建了其限制性图谱。当将pUE109导入大肠杆菌HB101时,其对紫外线的抗性并未增加。用pUE109转化耐辐射嗜放射菌未能检测到四环素抗性转化体。

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