Reduction of spontaneous mutation rates in cells
OwnerHeike Strobel, Wolfgang Schoenfeldinventors
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Abstract
The present invention relates to processes for reducing the spontaneous mutation frequencies in cells or organisms and for producing such cells and organisms, to cells and/or organisms with reduced spontaneous mutation frequencies and to processes for the generation of expression systems for proteins, for the production of proteins and for the production of fermentation products by using cells with reduced spontaneous mutation frequencies.
Claims62
Process for reducing the spontaneous mutation frequencies in a cell or an organism by introducing at least two mutations, whose combined actions lead to at least two enhanced cellular DNA repair mechanisms, into the cell or organism.
Process for producing a cell or an organism with reduced spontaneous mutation frequencies by introducing at least two mutations, whose combined actions lead to at least two enhanced cellular DNA repair mechanisms, into at least one cell of the organism and regenerating the organism therefrom if the organism is a multicellular organism.
E. coli MG1655dinB10 containing plasmid pmutL (DSM 17016).
E. coli MG1655dinB10 mutL::tet containing plasmid pmutL (DSM 17017).
E. coli MG1655 dnaE zae::cm containing plasmid pmutL (DSM 17018).
E. coli MG1655 dnaE zae::cm mutL::tet containing plasmid pmutL (DSM 17019).
E. coli MG1655dinB10 dnaE zae::cm (DSM 17015).
E. coli MG1655dinB10 dnaE zae::cm mutL::tet (DSM 17014).
E. coli MG1655dinB10 dnaE zae::cm containing plasmid pmutL (DSM 17020).
E. coli MG1655dinB10 dnaE zae::cm mutL::tet containing plasmid pmutL (DSM 17021).
Process for the generation of an expression system for a protein wherein the amino acid sequence of the protein is stabilized against spontaneously occurring mutations comprising:
- a) inserting a nucleic acid sequence encoding the protein into the genome of a host cell, that contains at least two mutations whose combined actions lead to an enhanced capability of at least two cellular DNA repair mechanisms to repair spontaneously occurring mutations, under the functional control of one or more regulatory units allowing an inducible or constitutive expression of the protein, or
- b) inserting a nucleic acid sequence encoding the protein into a vector under the functional control of one or more regulatory units allowing an inducible or constitutive expression of the protein and transferring the vector into a host cell, that contains at least two mutations whose combined actions lead to an enhanced capability of at least two cellular DNA repair mechanisms to repair spontaneously occurring mutations and
- c) culturing and/or maintaining the host cell in an appropriate medium.
Process for the production of a protein wherein the amino acid sequence of the protein is stabilized against spontaneously occurring mutations comprising:
- a) inserting a nucleic acid sequence encoding the protein into the genome of a host cell, that contains at least two mutations whose combined actions lead to an enhanced capability of at least two cellular DNA repair mechanisms to repair spontaneously occurring mutations, under the functional control of one or more regulatory units allowing an inducible or constitutive expression of the protein, or
- b) inserting a nucleic acid sequence encoding the protein into a vector under the functional control of one or more regulatory units allowing an inducible or constitutive expression of the protein and transferring the vector into a host cell, that contains at least two mutations whose combined actions lead to an enhanced capability of at least two cellular DNA repair mechanisms to repair spontaneously occurring mutations,
- c) culturing the host cell in an appropriate medium under conditions allowing the expression of the protein, and
- d) isolating the protein expressed.
Process for the production of a fermentation product by cultivating a cell producing the fermentation product and/or at least one enzyme involved in the formation of the fermentation product in a medium wherein the genome of the cell is stabilized against spontaneously occurring sequence changes by at least two mutations whose combined actions lead to an enhanced capability of at least two cellular DNA repair mechanisms to repair spontaneously occurring mutations.
Dependencies are inferred from the claim text.