Thermophilic DNA polymerases from thermoactinomyces vulgaris

Abstract

The present invention provides compositions comprising thermostable DNA polymerases derived from hyperthermophilic eubacteria. In particular, the present invention comprises thermostable DNA polymerases from the hyperthermophilic eubacterial species Thermoactinomyces vulgaris . The present invention also provides methods for utilizing naturally-occurring and non-naturally-occurring forms of T. vulgaris DNA polymerase in sequencing, reverse transcription, and amplification reactions.

1 - 38 . (canceled)  
     
     
         39 . A composition comprising a purified and isolated DNA polymerase comprising an amino acid sequence selected from the group consisting of sequences that are at least 99.8% identical to SEQ ID NO: 4 and sequences that are at least 99.8% identical to SEQ ID NO: 6.  
     
     
         40 . The composition of  claim 39 , wherein said polymerase is selected from the group consisting of SEQ NO:4 with a single amino acid substitution and SEQ ID NO:6 with a single amino acid substitution.  
     
     
         41 . The composition of  claim 39 , wherein said polymerase lacks 5′-3′ exonuclease activity.  
     
     
         42 . The composition of  claim 39 , wherein said polymerase has DNA synthesis activity.  
     
     
         43 . The composition of  claim 39 , wherein said polymerase has reverse transcriptase activity.  
     
     
         44 . The composition of  claim 39 , wherein said polymerase has a molecular weight of about 97 kD.  
     
     
         45 . The composition of  claim 43 , wherein said polymerase has reverse transcriptase activity in the presence of magnesium ions.  
     
     
         46 . The composition of  claim 45 , wherein said reverse transcriptase activity is substantially manganese ion independent.  
     
     
         47 . The composition of  claim 39 , wherein said DNA polymerase is thermostable.  
     
     
         48 . The composition of  claim 39 , wherein said composition is substantially free of endonuclease activity.  
     
     
         49 . The composition of  claim 39 , wherein said composition is greater than 85% pure.  
     
     
         50 . The composition of  claim 39 , wherein said composition is greater than 95% pure.  
     
     
         51 . The composition of  claim 39 , wherein said polymerase is a native polymerase.  
     
     
         52 . The composition of  claim 39 , wherein said polymerase is a recombinant polymerase.  
     
     
         53 . The composition of  claim 39 , wherein said polymerase comprises an amino acid sequence that is at least 99.8% identical to SEQ ID NO: 4.  
     
     
         54 . The composition of  claim 39 , wherein said polymerase comprises an amino acid sequence that is at least 99.8% identical to SEQ ID NO: 6.