Exonuclease deficient polymerases
Described herein are polymerase variants that are exonuclease deficient. Some variants retain the strand displacement capability comparable to the wild-type or parental polymerase. Some variants have a strand displacement capability that is improved relative to the wild-type or parental polymerase. The variants may have an extension rate that is greater than the wild-type or parental polymerase. The variants may have a waiting time that is less than the wild-type or parental polymerase.
1. A modified DNA polymerase, derived from a parental DNA polymerase polypeptide having a DNA polymerase activity comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 1, comprising a substitution at a position corresponding to a position of SEQ ID NO: 2, wherein the modified polymerase is exonuclease deficient.
2. The modified DNA polymerase according to claim 1 , wherein said modified polymerase retains a strand displacement capability.
3. The modified DNA polymerase according to claim 1 , wherein said modified polymerase has a substitution selected from the group consisting of Y242G, Y242A, Y242L, and Y242S.
4. The modified DNA polymerase according to claim 1 , wherein said modified polymerase has at least 90%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 1.
5. The modified DNA polymerase according to claim 1 , wherein said modified polymerase has an extension rate that is greater than the parental polymerase.
6. The modified DNA polymerase of claim 5 , wherein the extension rate is between 1.5 to 5 times greater than the parental polymerase.
7. The modified DNA polymerase of claim 5 , wherein the extension rate is at least 1.5 times greater than the parental polymerase.
8. The modified DNA polymerase according to claim 1 , wherein said modified polymerase has a median waiting time that is less than the wild-type or parental polymerase.
9. The modified DNA polymerase according to claim 8 , wherein said modified polymerase has a median waiting time that is less than the waiting time of the polymerase according to SEQ ID NO: 11.
10. The modified DNA polymerase according to claim 8 , wherein said modified polymerase has a median waiting time that is less than 3 seconds.
11. The modified DNA polymerase according to claim 1 , wherein said modified polymerase has a substitution corresponding to Y242G.
12. The modified DNA polymerase according to claim 1 , wherein said modified polymerase has a substitution corresponding to Y242A.
13. The modified DNA polymerase according to claim 1 , wherein said modified polymerase has a substitution corresponding to Y242L.
14. The modified DNA polymerase according to claim 1 , wherein said modified polymerase has a substitution corresponding to Y242S.
15. The modified DNA polymerase according to claim 1 , wherein said modified polymerase comprises T529M+S366A+A547F+Y242G/A/L/S.