Polypeptides having nucleic acid binding activity
View Patent ↗Polynucleotides having nucleic acid binding activity are provided. Methods of stabilizing a nucleic acid duplex are provided. Methods of promoting the annealing of complementary nucleic acid strands are provided. Methods of increasing the processivity of a DNA polymerase are provided. Methods of enhancing the activity of a nucleic acid modification enzyme are provided. Fusion proteins are provided. Methods of using fusion proteins are provided. Kits are provided.
1. A method of increasing the processivity of a DNA polymerase comprising fusing the DNA polymerase to a polypeptide having at least 95% identity to SEQ ID NO:1, wherein the polypeptide has nucleic acid binding activity.
2. A method of increasing the processivity of a DNA polymerase comprising combining a reaction mixture comprising the DNA polymerase with a polypeptide comprising an amino acid sequence having at least 95% identity to SEQ ID NO:1, wherein the polypeptide has nucleic acid binding activity.
3. The method of claim 1 or 2 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:1 or a fragment of SEQ ID NO:1 that has nucleic acid binding activity.
4. The method of claim 1 , wherein the polypeptide has double-stranded nucleic acid binding activity.
5. The method of claim 1 , wherein the DNA polymerase is thermostable.
6. The method of claim 1 , wherein the polypeptide has a basic isoelectric point.
7. The method of claim 1 , wherein the polypeptide promotes annealing of complementary nucleic acid strands.
8. The method of claim 1 , wherein the DNA polymerase lacks 5′-3′ exonuclease activity.
9. The method of claim 1 , wherein the DNA polymerase is selected from the group consisting of Taq polymerase, Tne polymerase, Tma polymerase, Tfi polymerase, Tth polymerase, and Pfu polymerase.
10. The method of claim 1 , wherein the DNA polymerase and the polypeptide are fused by a linker.
11. The method of claim 1 , wherein the processivity of the DNA polymerase is increased in a primer extension reaction.
12. The method of claim 11 , wherein the primer extension reaction is a polymerase chain reaction (PCR).
13. The method of claim 12 , wherein the PCR is hot start PCR.
14. The method of claim 2 , wherein the polypeptide has double-stranded nucleic acid binding activity.
15. The method of claim 2 , wherein the DNA polymerase is thermostable.
16. The method of claim 2 , wherein the polypeptide has a basic isoelectric point.
17. The method of claim 2 , wherein the polypeptide promotes annealing of complementary nucleic acid strands.
18. The method of claim 2 , wherein the DNA polymerase lacks 5′-3′ exonuclease activity.
19. The method of claim 2 , wherein the DNA polymerase is selected from the group consisting of Taq polymerase, Tne polymerase, Tma polymerase, Tfi polymerase, Tth polymerase, and Pfu polymerase.
20. The method of claim 2 , wherein the processivity of the DNA polymerase is increased in a primer extension reaction.
21. The method of claim 20 , wherein the primer extension reaction is a polymerase chain reaction (PCR).
22. The method of claim 21 , wherein the PCR is hot start PCR.