C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
195/1.14, 195/1.
C12N 15/70 (2006.01) C07K 14/485 (2006.01) C07K 14/56 (2006.01) C07K 14/65 (2006.01) C12N 15/69 (2006.01) C12N 15/73 (2006.01)
Patent
CA 1312026
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 opera- tive to: (1) confer upon the plasmid the capacity for autonomous replication in a host cell; (2) control auton- omous plasmid replication in relation to the temperature at which host cell cultures are maintained; (3) stabi- lize 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 endonuc- lease recognition sites, unique to the plasmid and faci- litative of exogenous gene DNA sequence insertion; and (6) terminate mRNA transcription of adjacent DNA sequen- ces 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 pro- vide a strong promoter of mRNA transcription function- ally 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).
460748
Amgen Inc.
Ridout & Maybee Llp
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