Cloning systems for Rhodococcus and related bacteria
Abstract
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.
- Inventors:
- Issue Date:
- OSTI Identifier:
- 7273216
- Patent Number(s):
- 4952500
- Application Number:
- PPN: US 7-151319
- Assignee:
- Univ. of Georgia Research Foundation, Inc., Athens, GA (United States)
- DOE Contract Number:
- AS09-80ER10683; FG09-86ER13588
- Resource Type:
- Patent
- Resource Relation:
- Patent File Date: 1 Feb 1988
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 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
Citation Formats
Finnerty, W R, and Singer, M E. Cloning systems for Rhodococcus and related bacteria. United States: N. p., 1990.
Web.
Finnerty, W R, & Singer, M E. Cloning systems for Rhodococcus and related bacteria. United States.
Finnerty, W R, and Singer, M E. Tue .
"Cloning systems for Rhodococcus and related bacteria". United States.
@article{osti_7273216,
title = {Cloning systems for Rhodococcus and related bacteria},
author = {Finnerty, W R and Singer, M E},
abstractNote = {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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1990},
month = {8}
}