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US 7,125,695 B2Granted
Directed evolution of microorganisms
AssigneeGenencor International, Inc.
InventorsVolker Schellenberger, Amy D. Liu, Olga V. Selifonova
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Abstract
The present invention provides methods for directing the evolution of microorganisms comprising the use of mutator genes and growth under conditions of selective pressure. The method discloses mutator genes which can be used in the methods of the present invention and provides ATCC deposits which exemplify the evolved microorganisms produced by the methods.
Claims20
A method for producing a protein from an evolved microorganism comprising the steps of:
- a) obtaining a microorganism comprising at least one heterologous mutator gene and at least one introduced nucleic acid encoding at least one heterologous protein;
- b) culturing said microorganism for at least 20 doublings under conditions suitable for selection of an evolved microorganism, wherein said heterologous mutator gene generates a mutation rate of at least 5–100,000 fold relative to wild type, and wherein said heterologous protein is expressed by said microorganism;
- and
- c) restoring said evolved microorganism to a wild type mutation rate.
1 more element
A method for producing a heterologous protein in an evolved microorganism comprising the steps of:
- a) obtaining a microorganism comprising at least one heterologous mutator gene and at least one introduced nucleic acid encoding at least one heterologous protein, wherein said at least one heterologous protein is an enzyme necessary for an enzymatic pathway in a microorganism;
- b) culturing said microorganism for at least 20 doublings under conditions suitable for selection of an evolved microorganism, wherein said heterologous mutator gene generates a mutation rate of at least 5 to 100,000-fold relative to wild type, and wherein said heterologous protein is expressed by said microorganism;
- and
- c) restoring said evolved microorganism to a wild type mutation rate.
1 more element
Dependencies are inferred from the claim text.