Method and a hybrid promoter for controlling exogenous gene transcription
AssigneeAmgen Inc.
InventorGrant A. Bitter
Life of this patent
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Abstract
A hybrid promoter for controlling exogenous gene transcription is constructed by insertion of a UAS.sub.G into a GPD portable promoter. The resulting hybrid promoter is placed upstream of an exogenous gene in a hybrid yeast-bacterial plasmid, which is used to transform yeast cells. Due to the regulation imparted to the GPD promoter by the UAS.sub.G, transcription of the exogenous gene, and hence production of the exogenous gene product, may be regulated by controlling the composition of the carbon source in the yeast culture medium. Specifically, glucose is used to repress transcription and galactose is used to induce transcription.
Claims18
A hybrid promoter which functions is control transcription of an exogenous gene in yeast comprising:
- a portion of a yeast glyceraldehyde-3-phosphate dehydrogenase promoter effective for promoting transcription;
- a TATA box homology adjacent a downstream end of the hybrid promoter;
- and
- an insert comprising an extrinsic upstream regulatory sequence at a location in said glyceraldehyde-3-phosphate dehydrogenase promoter upstream of said TATA box homology.
A DNA segment mediating regulatable transcription in yeast comprising:
- a hybrid promoter which functions to control transcription of an exogenous gene in yeast comprising:
- a portion of a yeast glyceraldehyde-3-phosphate dehydrogenase promoter effective for promoting transcription;
- a TATA box homology adjacent a downstream end of the hybrid promoter;
- an insert comprising a portion of any extrinsic upstream regulatory sequence at a location in said glyceraldehyde-3-phosphate dehydrogenase promoter upstream of said TATA box homology, said insert functioning to regulate transcription;
- and
- an exogenous gene downstream of said TATA box homology.
A method for controlling transcription of a gene in yeast comprising the steps of:
- inserting a portion of an extrinsic upstream activating sequence upstream of a TATA box homology in a yeast glyceraldehyde-3-phosphate dehydrogenase promoter, said portion functioning to regulate transcription;
- and
- introducing an exogenous gene downstream of the TATA box homology.
Dependencies are inferred from the claim text.