Partial characterization of a small, multiple-copy plasmid from Streptomyces espinosus and the derivation of a high copy-number deletion mutant |
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Authors: | J J Manis S K Highlander |
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Institution: | The Upjohn Company, Fermentation Research and Development, 7171 Portage Road, Kalamazoo, MI 49001 U.S.A. |
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Abstract: | An organism classified as Streptomyces espinosus was found to carry an approx. 9.2-kb plasmid. This plasmid, designated pUC6, has a copy number of 30-40 per host genome equivalent. Plasmid pUC1061, a copy-number mutant of pUC6, was isolated after in vitro deletion of an approx. 2.0-kb XhoI restriction fragment. Plasmid pUC1061 has a copy number of 500-600. Plasmid pUC1061 appears to be incompatible with pUC6 and will transform a pUC6-containing culture at a frequency of approx. 1%. The sizes, restriction maps and copy numbers of plasmids pUC6 and pUC1061 indicate these may be valuable vectors for gene cloning Streptomyces. |
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Keywords: | Recombinant DNA vectors transformation incompatibility restriction maps EtBr ethidium bromide kb kilobase(s) PEG polyethylene glycol SDS sodium dodecyl sulfate |
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