Expression vectors with modified ColE1 origin of replication for control of plasmid copy number

US6806066B2 · kind B2 · utility

Assignee

• Boehringer Ingelheim International GmbH · DE

Inventors

• Karl Bayer · Brooklyn, US
• Reingard Grabherr · Pressbaum, AT
• Erik Nilsson · Stockholm, SE
• Gerald Striedner · Wien, AT

Key dates

• Filing date: Oct 3, 2001
• Grant date: Oct 19, 2004
• Expiry date: Oct 3, 2021

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/70
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

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.