Polymerases

Abstract

Presented herein are methods and compositions for thermostable DNA polymerases that may be used to improve the PCR process and to improve the results obtained when using a thermostable DNA polymerase in other recombinant techniques such as DNA sequencing, nick-translation, and reverse transcription.

1 . A recombinant DNA polymerase comprising the amino acid sequence of any one of SEQ ID NOs: 1-66. 
     
     
         2 . A recombinant DNA polymerase comprising an amino acid sequence that is at least 90%, 95%, 99% identical to any one of SEQ ID NOs: 1-66, and which recombinant DNA polymerase exhibits polymerase activity. 
     
     
         3 . The recombinant DNA polymerase according to  claim 2 , wherein the polymerase further comprises an N-terminal leader peptide. 
     
     
         4 . The recombinant DNA polymerase according to  claim 3 , wherein the leader peptide comprises a His tag. 
     
     
         5 . The recombinant DNA polymerase according to  claim 4 , wherein the leader peptide comprises the amino acid sequence of SEQ ID NO: 72. 
     
     
         6 . A nucleic acid molecule encoding an altered polymerase as defined in  claim 2 . 
     
     
         7 . An expression vector comprising the nucleic acid molecule of  claim 6 . 
     
     
         8 . A host cell comprising the vector of  claim 7 . 
     
     
         9 . A method for incorporating nucleotides into DNA comprising allowing the following components to interact: (i) a polymerase according to  claim 2 , (ii) a DNA template; and (iii) a nucleotide solution. 
     
     
         10 . The recombinant DNA polymerase according to  claim 2 , wherein the polymerase exhibits increased fidelity compared to Vent DNA polymerase. 
     
     
         11 . The recombinant DNA polymerase according to  claim 2 , wherein the polymerase exhibits increased activity compared to Vent DNA polymerase. 
     
     
         12 . The recombinant DNA polymerase according to  claim 2 , wherein the polymerase exhibits increased activity and increased fidelity compared to Vent DNA polymerase. 
     
     
         13 . The recombinant DNA polymerase according to  claim 2 , wherein the polymerase exhibits increased activity and lower fidelity compared to Vent DNA polymerase. 
     
     
         14 . The recombinant DNA polymerase according to  claim 2 , wherein the polymerase exhibits increased fidelity and lower activity compared to Vent DNA polymerase. 
     
     
         15 . A recombinant DNA polymerase comprising the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, or 56. 
     
     
         16 - 17 . (canceled) 
     
     
         18 . The recombinant DNA polymerase according to  claim 15 , wherein the polymerase further comprises an N-terminal leader peptide. 
     
     
         19 . The recombinant DNA polymerase according to  claim 18 , wherein the leader peptide comprises a His tag. 
     
     
         20 . The recombinant DNA polymerase according to  claim 19 , wherein the leader peptide comprises the amino acid sequence of SEQ ID NO: 72.