Cloning plasmid for streptomyces
AssigneeThe Upjohn Company
InventorsJack J. Manis, Sarah K. Highlander
Life of this patent
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Abstract
Novel chemical compound plasmid pUC1061 obtained by deletion of .about.2.0 kilobases of DNA from the Streptomyces espinosus plasmid pUC6. This plasmid is useful as a cloning vehicle in recombinant DNA work. For example, using DNA methodology, a desired gene, for example, the glucose isomerase gene, can be inserted into the plasmid and the resulting plasmid can then be transformed into a suitable host microbe which, upon culturing, produces the desired glucose isomerase.
Claims5
Streptomyces espinosus (pUC1061) having the deposit accession number NRRL 12488.
Plasmid pUC1061, characterized as shown by the restriction map in FIG. 2 of the drawings and having the following additional restriction sites:
- Sal I (5-6 sites) and Sst I (2 sites);
- lacking of restriction sites for the following restriction endonucleases:
- Bam HI, Hind III, Kpn I, Hpa I, EcoRI, and XbaI;
- and, having a molecular length of ca. 7.2 kilobases.
A process for preparing plasmid pUC1061 which comprises:
- (a) digestion pUC6 DNA with restriction endonuclease XhoI to obtain fragmented linear plasmid DNA;
- and,
- (b) ligating the largest fragment of said plasmid DNA to obtain plasmid pUC1061.
A process for cloning plasmid pUC1061 into a suitable host of the genus Streptomyces which comprises:
- (a) digestion of pUC6 DNA with restriction endonuclease XhoI to obtain fragmented linear plasmid DNA;
- (b) ligating the largest fragment of said plasmid DNA to obtain plasmid pUC1061;
- and,
- (c) transforming said plasmid into said suitable host.
Dependencies are inferred from the claim text.