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US 10,590,400 B2Granted
DNA polymerases with increased 3′-mismatch discrimination
AssigneeRoch Molecular Systems, Inc.
InventorsKeith Bauer, Fred Reichert, Thomas W. Myers
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Abstract
Disclosed are mutant DNA polymerases having increased 3′-mismatch discrimination relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.
Claims15
A DNA polymerase having increased 3′-mismatch discrimination activity compared with a control DNA polymerase, wherein the DNA polymerase comprises a motif in the polymerase domain comprising
- E-R-M-A-F-X3-M-P-V-Q-G-T-A-A-D, wherein:
- X3 is any amino acid other than N (SEQ ID NO:10);
- wherein the DNA polymerase has at least 90% sequence identity to the amino acid sequence of SEQ ID NO:1, 2, 3, 4, 5, 6, 7, 36, 37, 38, 39, 40 or 41, and wherein the control DNA polymerase has the same amino acid sequence as the DNA polymerase except that the amino acid of the control DNA polymerase at position X3 is N.
Dependencies are inferred from the claim text.