Enzymes

Abstract

The invention relates to a nucleic acid polymerase capable of producing a non-DNA nucleotide polymer from a DNA nucleotide template, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at one or more residues of the thumb region, said residues selected from: amino acids 651 to 679 (patch 10A); wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue E664. In one embodiment said polymerase comprises the mutations Y409G and E664K. In one embodiment said polymerase comprises amino acid sequence corresponding to SEQ ID NO:12. The invention also relates to A nucleic acid polymerase capable of reverse transcribing a HNA nucleotide polymer into a DNA nucleotide polymer, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of sEQ ID NO:1, wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue I521.

We claim: 
     
         1 . A recombinant nucleic acid polymerase capable of producing a non-DNA nucleotide polymer from a DNA nucleotide polymer template, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated, relative to the amino acid sequence of SEQ ID NO:1, at one or more residues of the thumb region, said residues selected from amino acids 651 to 679 (patch 10A); and wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue E664. 
     
     
         2 . The recombinant nucleic acid polymerase of  claim 1 , wherein said non-DNA nucleotide polymer is a RNA polymer; and wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue Y409. 
     
     
         3 . The recombinant nucleic acid polymerase of  claim 2 , wherein said polymerase comprises the mutations Y409G and E664K. 
     
     
         4 . A method for making a RNA nucleotide polymer, said method comprising contacting a DNA template with a nucleic acid polymerase of  claim 2  and incubating to allow polymerization. 
     
     
         5 . A method for making a RNA nucleotide polymer, said method comprising contacting a DNA template with a nucleic acid polymerase of  claim 3  and incubating to allow polymerization. 
     
     
         6 . The recombinant nucleic acid polymerase of  claim 1 , wherein said polymerase is capable of producing a HNA nucleotide polymer from a DNA nucleotide polymer template and said polymerase comprises amino acid sequence corresponding to amino acids 651 to 679 (patch 10A) of SEQ ID NO:12. 
     
     
         7 . The recombinant nucleic acid polymerase of  claim 6 , wherein said polymerase is capable of producing a HNA nucleotide polymer from a DNA nucleotide polymer template and said polymerase comprises amino acid sequence corresponding to SEQ ID NO:12. 
     
     
         8 . A recombinant nucleic acid polymerase capable of reverse transcribing a HNA nucleotide polymer into a DNA nucleotide polymer, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated, relative to the amino acid sequence of SEQ ID NO:1, at residue I521. 
     
     
         9 . The recombinant nucleic acid polymerase according to  claim 8 , wherein said polymerase comprises a mutation selected from the group consisting of I521L, I521P and I521H. 
     
     
         10 . The recombinant nucleic acid polymerase of  claim 9 , wherein said polymerase comprises the mutation I521L. 
     
     
         11 . The recombinant nucleic acid polymerase of  claim 8 , wherein said polymerase further comprises the mutation A485L. 
     
     
         12 . The recombinant nucleic acid polymerase of  claim 11 , wherein said polymerase further comprises the mutation V93Q. 
     
     
         13 . The recombinant nucleic acid polymerase of  claim 12 , wherein said polymerase further comprises the mutations E141A and E143A. 
     
     
         14 . The recombinant nucleic acid polymerase of  claim 8 , wherein said non-DNA polymer is a HNA polymer and wherein said polymerase comprises the mutations I521L, A485L, V93Q, E141A, and E143A. 
     
     
         15 . A system comprising:
 (a) a recombinant nucleic acid polymerase capable of producing a non-DNA nucleotide polymer from a DNA nucleotide polymer template, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated, relative to the amino acid sequence of SEQ ID NO:1, at one or more residues of the thumb region, said residues selected from amino acids 651 to 679 (patch 10A); and wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue E664; and   (b) a recombinant nucleic acid polymerase capable of reverse transcribing a HNA nucleotide polymer into a DNA nucleotide polymer, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated, relative to the amino acid sequence of SEQ ID NO:1, at residue I521.