Cloning systems for Rhodococcus and related bacteria
A plasmid transformation system for Rhodococcus was developed using an Escherichia coli-Rhodococcus shuttle plasmid. Rhodococcus sp. H13-A contains three cryptic indigenous plasmids, designated pMVS100, pMVS200 and pMVS300, of 75, 19.5 and 13.4 kilobases (Kb), respectively. A 3.8 Kb restriction fragment of pMVS300 was cloned into pIJ30, a 6.3 Kb pBR322 derivative, containing the E. coli origin of replication (ori) and ampicillin resistance determinant (bla) as well as a Streptomyces gene for thiostrepton resistance, tsr. The resulting 10.1 Kb recombinant plasmid, designated pMVS301, was isolated from E. coli DH1 (pMVS301) and transformed into Rhodococcus sp. AS-50, a derivative of strain H13-A, by polyethylene glycol-assisted transformation of Rhodococcus protoplasts and selection for thiostrepton-resistant transformants. This strain was deposited with the ATCC on Feb. 1, 1988 and assigned ATCC 53719. The plasmid contains the Rhodococcus origin of replication. The plasmid and derivatives thereof can therefore be used to introduce nucleic acid sequences to and from Rhodococcus for subsequent expression and translation into protein. The isolated origin of replication can also be used in the construction of new vectors. 2 figs.
- DOE Contract Number:
- AS09-80ER10683; FG09-86ER13588
- Assignee:
- Univ. of Georgia Research Foundation, Inc., Athens, GA (United States)
- Patent Number(s):
- US 4952500; A
- Application Number:
- PPN: US 7-151319
- OSTI ID:
- 7273216
- Resource Relation:
- Patent File Date: 1 Feb 1988
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PLASMIDS
BIOCHEMICAL REACTION KINETICS
RHODOCOCCUS
CLONING
GENETIC ENGINEERING
AMINO ACID SEQUENCE
DISEASE RESISTANCE
GENE REGULATION
BACTERIA
BIOTECHNOLOGY
CELL CONSTITUENTS
KINETICS
MICROORGANISMS
MOLECULAR STRUCTURE
REACTION KINETICS
SULFUR-OXIDIZING BACTERIA
550200* - Biochemistry