Cells and method for producing proteins comprising an unconventional amino acid

US7736899B1 · kind B1 · utility

Assignee

• INSTITUT PASTEUR · FR

Inventors

• Philippe Marliere · Paris, FR
• Volker Doring · Stuttgart, DE
• Henning Mootz · Marburg, DE

Key dates

• Filing date: Oct 28, 1999
• Grant date: Jun 15, 2010
• Expiry date: Oct 28, 2019

Classification

• Technology area (CPC A): Human Necessities
• CPC primary: A61K48/00
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

The present invention relates to a method for providing bacterial or yeast cells with the capacity to produce a protein, the amino acid sequence of which comprises at least one unconventional amino acid. The method involves (a) introducing at least one missense mutation in a target codon of a gene encoding a protein required for the growth of the bacterial or yeast cells, where the mutated protein synthesized from the mutated gene is not functional in the bacterial or yeast cells. The method also involves (b) selecting the bacterial or yeast cells obtained in (a) in a culture medium which (1) does not contain a nutrient compensating for the loss of functionality of the mutated protein and (2) contains an unconventional amino acid which restores the functionality of the protein required for growth of the bacterial or yeast cells, in which the unconventional amino acid is that encoded by the target codon. The method also involves culturing the bacterial or yeast cells obtained in (b) in a culture medium containing the amino acid encoded by the target codon.