Induction quiescence in bacterial cells and uses thereof

C - Chemistry – Metallurgy – 12 – N

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C12N 15/67 (2006.01) C07K 14/195 (2006.01) C12N 1/21 (2006.01) C12N 15/69 (2006.01) C12N 15/70 (2006.01) C12N 15/74 (2006.01) C12P 21/02 (2006.01) C12Q 1/68 (2006.01) G01N 33/68 (2006.01)

Patent

CA 2248579

Rcd expression from an extra-chromosomal vector in bacterial cells in broth culture induces quiescence. Expression continues from extra-chromosomal vectors within the cells during quiescence, while chromosomal expression is suppressed. The cells may be used as factories for the production of large amounts of polypeptides of interest, particularly polypeptides which normally have an adverse effect on cell viability or growth. Expression from an extra- chromosomal vector of interest may be monitored, in view of the reduced background expression from the chromosome. Vector copy number may be amplified. Cell cycles may be synchronised. Suitable cells for rcd-induced quiescence include hns- strains of bacteria such as E. coli.

L'expression de rcd par un vecteur extrachromosomique dans les cellules bactériennes d'un bouillon de culture provoque la quiescence. L'expression par le vecteur extrachromosomique se poursuit dans les cellules pendant la quiescence tandis que l'expression chromosomique cesse. Les cellules peuvent servir d'usines destinées à produire d'importantes quantités de polypeptides désirés, et en particulier de polypeptides ayant normalement un effet adverse sur la viabilité ou la croissance des cellules. L'expression d'un vecteur extrachromosomique désiré peut être surveillée pour qu'il soit possible de réduire l'expression d'arrière plan des chromosomes. Le nombre de copies du vecteur peut être accru. Les cycles des cellules peuvent être synchronisés. Les cellules susceptibles de produire la quiescence induite par le rcd comportent celles de la souche hns?-¿ de bactéries telles que E. coli.

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