Expression vectors with modified cole1 origin of replication...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/70 (2006.01) C12N 1/21 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2424624

In an expression vector having a ColE1 replication system, the homology of the RNAI and RNAII of the ColE1 origin of replication to uncharged tRNAs is modified mutations in the coding region of the RNAI gene and corresponding mutations in the RNAII gene. The mutation results in one or more base exchanges in loop 1 and/or loop 2 and/or loop 3 of RNAI and RNAII. In methods using this vector for producing recombinant proteins, plasmid copy number is stably maintained. In methods for plasmid production, high plasmid copy numbers can be obtained.

Dans un vecteur d'expression à système de réplication ColE1, l'homologie de RNAI et RNAII de l'origine de réplication ColE1 par rapport à des ARNt non chargés est modifiée par des mutations dans la région codante du gène RNAI et par des mutations correspondantes dans le gène RNAII. La mutation résulte d'un ou plusieurs échanges de base dans la boucle 1 et/ou la boucle 2 et/ou la boucle 3 de RNAI et RNAII. Dans des procédés utilisant ce vecteur pour produire des protéines de recombinaison, le nombre de copies de plasmides est stabilisé. Dans des procédés de production de plasmides, il est possible d'obtenir un nombre important de copies de plasmides.

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