Mutant polymerases with fast elongating activity

Abstract

An isolated polypeptide having fast elongating polymerase activity. Also provided are kits containing the isolated polypeptide and isolated polynucleotides encoding the isolated polypeptide.

What is claimed is: 
     
         1 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:
 (i) SEQ ID NO: 22 or a variant thereof having at least 95% sequence identity to SEQ ID NO: 22, wherein position 430 of the variant of SEQ ID NO: 22 is serine (S), tryptophan (W), leucine (L), and the variant of SEQ ID NO: 22 has fast elongating polymerase activity; or   (ii) a fragment of (i) comprising position 430 of SEQ ID NO: 22 or serine (S), tryptophan (W), or leucine (L) at position 430 of the variant of SEQ ID NO: 22, wherein the fragment has fast elongating polymerase activity.   
     
     
         2 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises the variant having at least 95% sequence identity to SEQ ID NO: 22, position 430 of the variant of SEQ ID NO: 22 is serine (S), and the variant of SEQ ID NO: 22 has fast elongating polymerase activity. 
     
     
         3 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises the variant having at least 95% sequence identity to SEQ ID NO: 22, position 430 of the variant of SEQ ID NO: 22 is tryptophan (W), and the variant of SEQ ID NO: 22 has fast elongating polymerase activity. 
     
     
         4 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises the variant having at least 95% sequence identity to SEQ ID NO: 22, position 430 of the variant of SEQ ID NO: 22 is leucine (L), and the variant of SEQ ID NO: 22 has fast elongating polymerase activity. 
     
     
         5 . The isolated polypeptide of  claim 1 , wherein:
 the variant of SEQ ID NO: 22 comprises at least one of:
 position 331 of the variant of SEQ ID NO: 22 is proline (P); 
 position 348 of the variant of SEQ ID NO: 22 is lysine (K); 
 position 371 of the variant of SEQ ID NO: 22 is isoleucine (I); and 
 position 429 of the variant of SEQ ID NO: 22 is leucine (L); and 
   the variant of SEQ ID NO: 22 has fast elongating polymerase activity.   
     
     
         6 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises:
 (i) the variant having at least 95% sequence identity to SEQ ID NO: 22, position   position 348 of the variant of SEQ ID NO: 22 is lysine (K), position 429 of the variant of SEQ ID NO: 22 is leucine (L), and position 430 of the variant of SEQ ID NO: 22 is tryptophan (W), and the variant of SEQ ID NO: 22 has fast elongating polymerase activity; or   (ii) SEQ ID NO: 6 (KT-7).   
     
     
         7 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises:
 (i) the variant having at least 95% sequence identity to SEQ ID NO: 22, position 348 of the variant of SEQ ID NO: 22 is lysine (K), position 371 of the variant of SEQ ID NO: 22 is isoleucine (I), position 429 of the variant of SEQ ID NO: 22 is leucine (L), and position 430 of the variant of SEQ ID NO: 22 is serine (S), and the variant of SEQ ID NO: 22 has fast elongating polymerase activity; or   (ii) SEQ ID NO: 22 (KT-11).   
     
     
         8 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises:
 (i) the variant having at least 95% sequence identity to SEQ ID NO: 22, position 331 of the variant of SEQ ID NO: 22 is proline (P), position 348 of the variant of SEQ ID NO: 22 is lysine (K), position 429 of the variant of SEQ ID NO: 22 is leucine (L), and position 430 of the variant of SEQ ID NO: 22 is leucine (L), and the variant of SEQ ID NO: 22 has fast elongating polymerase activity; or   (ii) SEQ ID NO: 24 (KT-12).   
     
     
         9 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises:
 (i) the variant having at least 95% sequence identity to SEQ ID NO: 22, position 331 of the variant of SEQ ID NO: 22 is proline (P), position 348 of the variant of SEQ ID NO: 22 is lysine (K), position 429 of the variant of SEQ ID NO: 22 is leucine (L), and position 430 of the variant of SEQ ID NO: 22 is leucine (L), and the variant of SEQ ID NO: 22 has fast elongating polymerase activity; or   (ii) SEQ ID NO: 30 (FL-12).   
     
     
         10 . The isolated polypeptide of  claim 1 , wherein the isolated polypeptide comprises SEQ ID NO: 6 (KT-7); SEQ ID NO: 22 (KT-11); SEQ ID NO: 24 (KT-12); or SEQ ID NO: 30 (FL-12), and the isolated polypeptide has fast elongating polymerase activity. 
     
     
         11 . An isolated polynucleotide encoding the isolated polypeptide of  claim 1 . 
     
     
         12 . The isolated polynucleotide of  claim 11 , wherein the polynucleotide comprises a nucleic acid sequence of SEQ ID NO: 5 (KT-7), SEQ ID NO: 21 (KT-11), SEQ ID NO: 23 (KT-12), or SEQ ID NO: 29 (FL-12). 
     
     
         13 . A kit for performing PCR assays, wherein the kit comprises an isolated polypeptide according to  claim 1 .