DNA plasmids

Details DNA plasmids DNA plasmids

1984-08-06

1987-12-01

Tanenholtz, Alvin E.

Chemistry: molecular biology and microbiology

Vector, per se

4351723, 935 29, 935 41, 935 42, C12N 100, C12N 1500

Patent

active

047104732

ABSTRACT:
Disclosed are novel circular DNA plasmids useful as vectors in recombinant methods to secure high levels of E.coli expression of exogenous genes. Plasmids of the invention comprise discrete DNA sequences operative to: (1) confer upon the plasmid the capacity for autonomous replication in a host cell; (2) control autonomous plasmid replication in relation to the temperature at which host cell cultures are maintained; (3) stabilize maintenance of the plasmid in host cell populations; (4) direct synthesis of a protein product indicative of plasmid maintenance in a host cell population; (5) provide, in series, a plurality of restriction endonuclease recognition sites, unique to the plasmid and facilitative of exogenous gene DNA sequence insertion; and (6) terminate mRNA transcription of adjacent DNA sequences and situated so as to terminate transcription of exogenous gene sequences inserted within the plasmid at said unique restriction endonuclease restriction sites. Plasmids preferably have a size of less than 5.0 kilobases (exclusive of any inserted exogenous gene) and optionally include a DNA sequence operative to provide a strong promoter of mRNA transcription functionally associated with a temperature sensitive repressor sequence. A presently preferred embodiment of novel plasmids of the invention is plasmid pCFM414 (A.T.C.C. No. 40076).

REFERENCES:
Light et al., EMBO J., vol. 2, pp. 93-98, 1983.
Rosen et al., Molec. Gen Genet., vol. 179, pp. 527-537 (1980).
Rao et al., Gene, vol. 3, pp. 247-263, 1978.
Bittner et al., Gene, vol. 15, pp. 319-329, (1981).
Meacock et al., Cell, vol. 20, pp. 529-542, (1980).
Derynck et al., Nature, vol. 287, pp. 193-197.
Rao et al., Gene, vol. 7, pp. 79-82 (1979).
Gentz et al., PNAS USA, vol. 78, pp. 4936-4940, Aug. 1981.

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