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US 6,605,431 B1Granted
Promoter elements and methods of use
AssigneeWisconsin Alumni Research Foundation
InventorsRichard L. Gourse, Shawn T. Estrem, Wilma E. Ross, Tamas Gaal
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Abstract
The present invention provides novel polynucleotides that include promoter elements. The present invention also provides methods and kits for identification of compounds that alter transcription, preferably decrease transcription, of a polynucleotide. Also provided by the present invention are methods directed to producing RNA polynucleotides and polypeptides.
Claims12
A method for detecting whether a compound alters transcription of a transcription unit, the method comprising:
- providing in a reaction mixture a first polynucleotide comprising a first promoter operably linked to a transcription unit, wherein the first promoter comprises an accessory promoter element and a core promoter element, wherein the accessory promoter element is selected from the group consisting of a distal accessory promoter element, a proximal accessory promoter element, and a combined accessory promoter element, and wherein the reaction mixture comprises at least one RNA polymerase;
- adding an amount of the compound to be tested to the reaction mixture under conditions effective to cause transcription;
- detecting an amount of a transcription product;
- and
- comparing the amount of the transcription product in the presence of the compound to an amount of the transcription product in the absence of the compound under the same conditions, wherein the compound does not alter the amount of a transcription product from a second polynucleotide, the second polynucleotide comprising a second promoter operably linked to the same transcription unit that is operably linked to the first promoter, the second promoter consisting of the same core promoter element of the first promoter.
2 more elements
Dependencies are inferred from the claim text.