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US 7,390,636 B2Granted
Method for construction and use of Kluyveromyces lactis promoter variants in K. lactis that substantially lack E. coli transcriptional capability
AssigneeNew England Biolabs, Inc.
InventorsChristopher Taron, Paul Colussi
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Abstract
Methods and compositions are provided relating to production of recombinant protein in yeast. A modified PLAC4 is described where one or more mutations may be introduced into the Pribnow box-like sequences in the promoter. The modified promoter when placed upstream of a target gene in a vector causes a significant reduction of target gene expression in transformed bacteria but produces efficient expression of the target gene in yeast.
Claims16
A method of producing a recombinant protein in yeast cells, comprising:
- (a) obtaining a vector into which a gene encoding the target protein has been inserted, the vector further comprising:
- a modified PLAC4 having a mutation in one or more Pribnow box-like sequences wherein the modification results in a significant reduction in gene expression when the vector is cloned in bacteria;
- (b) transforming yeast cells with the vector;
- and
- (c) producing an effective amount of the recombinant protein in the yeast cells.
2 more elements
A vector, comprising:
- a modified PLAC4 promoter having a mutation in one or more Pribnow box-like sequences operatively linked to a gene encoding a target protein.
Dependencies are inferred from the claim text.