# US 4,393,137 A · Cloning plasmid for streptomyces

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- Number: US 4,393,137 A
- Document ID: US4393137A
- Status: Granted
- Kind: A
- Application number: 06/276,425
- Filed: 1981-06-22
- Granted: 1983-07-12
- Earliest claimed: 1981-06-22
- Assignees: The Upjohn Company
- Inventors: Jack J. Manis; Sarah K. Highlander
- Web page: https://patentgrep.com/patent/US4393137A

## CPC

- C12N 15/76: for Actinomyces; for Streptomyces
- Y10S 435/886: Streptomyces
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## 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.

## Claims

### Claim 1

Streptomyces espinosus (pUC1061) having the deposit accession number NRRL 12488.

### Claim 2

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.

### Claim 3

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.

### Claim 4

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.

### Claim 5 (depends on 4)

A process, according to claim 4, wherein said Streptomyces is S. espinosus biotype 23724a.