US2007059713A1PendingUtilityA1

SSB-DNA polymerase fusion proteins

52
Individually held — no corporate assignee on recordPriority: Sep 9, 2005Filed: Sep 9, 2005Published: Mar 15, 2007
Est. expirySep 9, 2025(expired)· nominal 20-yr term from priority
Inventors:Jun LeeRobert PotterDavid Mandelman
C12N 15/62C12N 9/1276C12Q 1/6848C12Q 1/6806C12N 9/1252
52
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Claims

Abstract

Fusion proteins comprising a single strand DNA binding protein and a nucleic acid polymerase (e.g. DNA polymerase or reverse transcriptase). These high fidelity proteins are suitable for use in nucleic acid amplification methods, including the polymerase chain reaction (PCR).

Claims

exact text as granted — not AI-modified
1 . An isolated or purified single stranded binding protein (SSB)-nucleic acid polymerase fusion protein.  
     
     
         2 . The fusion protein of  claim 1  wherein said nucleic acid polymerase is a DNA polymerase.  
     
     
         3 . The fusion protein of  claim 1 , wherein said nucleic acid polymerase is a reverse transcriptase.  
     
     
         4 . The fusion protein of  claim 1 , wherein said nucleic acid polymerase is a thermostable DNA polymerase.  
     
     
         5 . The fusion protein of  claim 4 , wherein said thermostable DNA polymerase is  Thermococcus zilligi  (Tzi) DNA polymerase.  
     
     
         6 . The fusion protein of  claim 1 , wherein said SSB is a thermostable SSB.  
     
     
         7 . The fusion protein of  claim 6 , wherein said SSB is  Sulfolobus solfataricus  (Sso) SSB.  
     
     
         8 . A nucleic acid molecule encoding an SSB-nucleic acid polymerase fusion protein.  
     
     
         9 . The nucleic acid molecule of  claim 8  wherein said nucleic acid polymerase is a DNA polymerase.  
     
     
         10 . The nucleic acid molecule of  claim 8  wherein the nucleic acid polymerase is a reverse transcriptase.  
     
     
         11 . The nucleic acid molecule of  claim 9  wherein the DNA polymerase is a thermostable DNA polymerase.  
     
     
         12 . The nucleic acid molecule of  claim 11  wherein the thermostable DNA polymerase is  Thermococcus zilligi  (Tzi) DNA polymerase.  
     
     
         13 . The nucleic acid molecule of  claim 8  wherein said SSB is a thermostable SSB.  
     
     
         14 . The nucleic acid molecule of  claim 13  wherein said SSB is  Sulfolobus solfataricus  (Sso) SSB.  
     
     
         15 . An expression vector comprising the nucleic acid sequence of  claim 8  operably linked to a heterologous promoter.  
     
     
         16 . A host cell comprising the expression vector of  claim 15 .  
     
     
         17 . A nucleic acid molecule encoding a  Sulfolobus solfataricus  (Sso) SSB- Thermococcus zilligi  (Tzi) DNA polymerase fusion protein.  
     
     
         18 . A method of increasing the yield of a polymerase reaction on a target nucleic acid comprising: 
 (a) contacting a target-nucleic acid molecule with a primer which specifically hybridizes thereto, and an isolated or purified single-stranded binding protein (SSB)-nucleic acid polymerase fusion protein which binds to single stranded DNA and enhances the processivity of said nucleic acid polymerase compared to said polymerase which does not have a SSB fused thereto; and    (b) incubating the resulting mixture under conditions whereby said primer is extended by said polymerase.    
     
     
         19 . The method of  claim 18  wherein said nucleic acid polymerase is a DNA polymerase.  
     
     
         20 . The method of  claim 18  wherein said nucleic acid polymerase is a reverse transcriptase.  
     
     
         21 . The method of  claim 18  wherein said DNA polymerase is a thermostable DNA polymerase.  
     
     
         22 . The method of  claim 21  wherein said DNA polymerase is  Thermococcus zilligi  DNA polymerase.  
     
     
         23 . The method of  claim 18 , wherein said SSB is a thermostable SSB.  
     
     
         24 . The method of  claim 23  wherein said thermostable SSB is  Sulfolobus solfataricus  SSB.  
     
     
         25 . An isolated or purified  Sulfolobus solfataricus  (Sso) SSB- Thermococcus zilligi  (Tzi) DNA polymerase fusion protein

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