Chromosomal expression of heterologous genes in bacterial cells

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/90 (2006.01) C07K 14/395 (2006.01) C07K 14/47 (2006.01) C07K 14/495 (2006.01) C07K 14/65 (2006.01) C12N 9/02 (2006.01) C12N 9/16 (2006.01) C12N 15/66 (2006.01) C12N 15/67 (2006.01) C12N 15/70 (2006.01) C12N 15/74 (2006.01)

Patent

CA 2179656

The present invention provides compositions and methods for inserting a copy of a heterologous gene into the chromosome of a host cell such as a E. coli through the use of a chromosomal transfer DNA, a circular, non-self replicating DNA carrying a site-specific recombination site such as lambda att P. The host cell chromosome contains a second site-specific recombination site. When the chromosomal transfer DNA is introduced into the host cell, expression of an integration enzyme such as integrase causes the integration of the chromosomal transfer DNA into the host cell chromosome at the second recombination site.

L'invention concerne des compositions et des procédés d'insertion d'une copie d'un gène hétérologue dans le chromosome d'une cellule hôte telle que <u>E. coli</u>, à l'aide d'un ADN de transfert chromosomique, à un ADN circulaire ne se repliquant pas automatiquement, porteur d'un site de recombinaison à spécificité de sites tel que <u>att</u> P lambda. Le chromosome de la cellule hôte contient un second site de recombinaison à spécificité de site. Lorsque l'on introduit l'ADN de transfert chromosomique dans la cellule hôte, l'expression d'une enzyme d'intégration telle que l'intégrase provoque l'intégration de l'ADN de transfert chromosomique dans le chromosome de la cellule hôte sur le second site de recombinaison.

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