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The plasmids of Deinococcus spp. and the cloning and restriction mapping of the D. radiophilus plasmid pUE1
Authors:Martin W. Mackay  Ghalib H. Al-Bakri  B. E. B. Moseley
Affiliation:(1) Department of Microbiology, University of Edinburgh, School of Agriculture, EH9 3JG Edinburgh, UK;(2) Present address: Department of Molecular Biology, University of Edinburgh, Mayfield Road, EH9 3JR Edinburgh, UK
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.
Keywords:Cloning  Deinococcus radiophilus  Deinococcus spp.  Plasmids  Restriction mapping
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