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 nucleic acid encoding a fusion polypeptide comprising a sequence non-specific double-stranded nucleic acid binding domain that has at least 75% identity to SEQ ID NO:2, joined to a reverse transcriptase.
2. The nucleic acid of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding domain has at least 85% identity to SEQ ID NO:2.
3. The nucleic acid of claim 1 , wherein the sequence non-specific double-stranded nucleic acid binding domain has at least 95% identity to SEQ ID NO:2.
4. The nucleic acid of 1 , wherein the sequence non-specific double-stranded nucleic acid binding domain comprises the amino acid sequence of SEQ ID NO:2.
5. An expression vector comprising a nucleic acid of claim 1 .
6. A host cell comprising a nucleic acid encoding a fusion polypeptide wherein the fusion polypeptide comprises a sequence non-specific double-stranded nucleic acid binding domain that has at least 75% identity to SEQ ID NO:2, joined to a reverse transcriptase.
7. The host cell of claim 6 , wherein the sequence non-specific double-stranded nucleic acid binding domain has at least 85% identity to SEQ ID NO:2.
8. The host cell of claim 6 , wherein the sequence non-specific double-stranded nucleic acid binding domain has at least 95% identity to SEQ ID NO:2.
9. The host cell of claim 6 , wherein the sequence non-specific double-stranded nucleic acid binding domain comprises the amino acid sequence of SEQ ID NO:2.
10. A method of producing a fusion polypeptide comprising a sequence non-specific double-stranded nucleic acid binding domain that has at least 75% identity to SEQ ID NO:2 joined to a reverse transcriptase, the method comprising culturing a host cell of claim 6 under conditions in which the fusion polypeptide is expressed.
11. The method of claim 10 , further comprising purifying the fusion polypeptide produced by the host cell.