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US 6,043,038 AGranted
High-throughput screening assays for modulators of primase activity
AssigneeTularik Inc.
InventorsMohanram Sivaraja, Ashok Sanadi, M. Gregory Peterson
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Abstract
This invention provides novel assays for primase activity and for identifying modulators of primase activity. The assays include both solid-phase and liquid-phase methods that are amenable to high throughput screening methods. The assays of the invention are readily adaptable to numerous types of primase, and are capable of identifying novel classes of primase activity modulators.
Claims28
A method for measuring the activity of a primase, the method comprising the steps of:
- incubating a mixture comprising a DNA template, a primase, and ribonucleotide triphosphates under conditions in which the primase polymerizes the triphosphates on the template to form a nucleic acid comprising one or more DNA-RNA heterohybrid regions;
- contacting the nucleic acid with a recognition reagent that comprises a tag and a moiety which specifically binds to a DNA-RNA heterohybrid region;
- binding the tag to a solid support;
- and
- detecting the presence of DNA-RNA heterohybrid regions immobilized on the solid support.
2 more elements
A method for measuring the activity of a primase, the method comprising the steps of:
- providing a DNA template immobilized on a solid support;
- contacting the DNA template with a reaction mixture comprising a primase and ribonucleotide triphosphates under conditions in which the primase polymerizes the triphosphates on the template to form a nucleic acid comprising one or more DNA-RNA heterohybrid regions;
- and
- detecting the presence of DNA-RNA heterohybrid regions bound to the solid support.
1 more element
Dependencies are inferred from the claim text.