Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1988-02-01
1990-08-28
Wiseman, Thomas G.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
4351723, 4352523, 435320, 435863, 435826, 435872, 435886, 435 91, 536 27, 935 6, 935 22, 935 60, 935 72, C12P 2100, C12N 1500, C12N 122, C12R 132, C12R 104, C12R 1365, C12R 1465, C07H 1512
Patent
active
049525000
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.
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Singer, M. E., 1986, Abstracts of the Annual Meeting of the American Society for Microbiology, Mar. 23-28, 1986, Abstract No. H-88, "Plasmid Transformation in Arthrobacter sp H13A", p. 142, col. 1.
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Singer, et al., "Microbial Desulfurization and Biosurfactant Production Strain Improvement Through Genetic Approaches", Fif. Intl. Symp. Gen. Indust. Microorg., M. Alacevice, et al., eds. (May 1987).
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Finnerty William R.
Singer Mary E.
Peet Richard C.
University of Georgia Research Foundation Inc.
Wiseman Thomas G.
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