Dna polymerases with increased 3'-mismatch discrimination
AssigneeRoche Molecular Systems, Inc.
InventorsFred Reichert, Keith Bauer, Thomas W. Myers, Nancy J. Schoenbrunner, Joseph San Filippo
Life of this patent
Estimating the term…
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.
Claims21
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
- A-G-X1-P-F-N-X4-N-S-X7-X8-Q-X9-X10-R-X12-L-F-X14-X15-L, wherein:
- X1 is H or E;
- X4 is L or I;
- X7 is R or P;
- X8 is D or K;
- X9 is L or V;
- X10 is any amino acid other than E or S;
- X12 is V or I;
- X14 is D or E;
- and
- X15 is E or K (SEQ ID NO:44).
The DNA polymerase of claim 11, wherein the DNA polymerase comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:
- 2.
Dependencies are inferred from the claim text.