Nucleic acid modifying enzymes
This invention provides for an improved generation of novel nucleic acid modifying enzymes. The improvement is the fusion of a sequence-non-specific nucleic-acid-binding domain to the enzyme in a manner that enhances the ability of the enzyme to bind and catalytically modify the nucleic acid.
1. A method of synthesizing DNA from an RNA template, the method comprising contacting the RNA template with a primer and a polypeptide comprising a sequence non-specific double-stranded nucleic acid binding polypeptide that has at least 75% identity to SEQ ID NO:2 or amino acids 7-71 of SEQ ID NO:10 joined to reverse transcriptase; and incubating the solution under conditions where the primer is extended.
2. The method of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide comprises the amino acid sequence of SEQ ID NO:2 or amino acids 7-71 of SEQ ID NO:10.
3. The method of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide has at least 80% identity to SEQ ID NO:2.
4. The method of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide comprises SEQ ID NO:2.
5. A polypeptide comprising a sequence non-specific double-stranded nucleic acid binding polypeptide that has at least 75% identity to SEQ ID NO:2, or amino acids 7-71 of SEQ ID NO:10, joined to reverse transcriptase.
6. The polypeptide of claim 5 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide comprises the amino acid sequence of SEQ ID NO:2 or amino acids 7-71 of SEQ ID NO:10.
7. The polypeptide of claim 5 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide has at least 80% identity to SEQ ID NO:2.
8. The polypeptide of claim 5 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide has at least 90% identity to SEQ ID NO:2.
9. The polypeptide of claim 5 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide comprises the amino acid sequence of SEQ ID NO:2.
10. The polypeptide of claim 5 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide has at least 80% identity to amino acids 7-71 of SEQ ID NO:10.
11. The polypeptide of claim 5 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide has at least 90% identity to amino acids7-71 of SEQ ID NO:10.
12. The polypeptide of claim 5 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide comprises amino acids 7-71 of SEQ ID NO:10.
13. The method of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide has at least 90% identity to SEQ ID NO:2.
14. The method of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide has at least 80% identity to amino acids 7-71 of SEQ ID NO:10.
15. The method of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide has at least 90% identity to amino acids 7-71 of SEQ ID NO:10.
16. The method of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding polypeptide comprises amino acids 7-71 of SEQ ID NO:10.