Multimers of S. solfataricus single-stranded DNA-binding protein and methods of use thereof

Abstract

The invention provides multimers of S. solfataricus ssDNA binding protein that bind single stranded DNA. The multimers are robust and stable reagents for use in PCR and other techniques for engineering DNA. The invention further provides methods for performing nucleic acid amplification and engineering using the multimers.

What is claimed is:  
     
         1 . An isolated multimer, wherein each unit of said multimer has at least 70% sequence identity to SEQ ID NO:1, and wherein said multimer binds single stranded DNA.  
     
     
         2 . An isolated multimer of  claim 1 , wherein each unit of said multimer has at least 80% sequence identity to SEQ ID NO:1.  
     
     
         3 . An isolated multimer of  claim 1 , wherein each unit of said multimer has at least 90% sequence identity to SEQ ID NO:1.  
     
     
         4 . An isolated multimer of  claim 1 , wherein each unit has the sequence of SEQ ID NO:1.  
     
     
         5 . An isolated multimer of  claim 1 , wherein said multimer is a tetramer.  
     
     
         6 . A method of performing nucleic acid amplification, said method comprising contacting a single stranded DNA with a multimeric protein, wherein each unit of said multimeric protein has at least 70% sequence identity to SEQ ID NO:1, and wherein said multimeric protein binds single stranded DNA.  
     
     
         7 . A method of  claim 6 , wherein each unit of said multimeric protein has at least 80% sequence identity to SEQ ID NO:1.  
     
     
         8 . A method of  claim 6 , wherein each unit of said multimeric protein has at least 90% sequence identity to SEQ ID NO:1.  
     
     
         9 . A method of  claim 6 , wherein each unit of said multimeric protein has the sequence of SEQ ID NO:1.  
     
     
         10 . A method of  claim 6 , wherein said nucleic acid amplification is selected from the group consisting of polymerase chain reaction, ligase chain reaction, transcription-based amplification system, and self-sustained sequence replication system.  
     
     
         11 . A method of  claim 10 , wherein the method of nucleic acid amplification is polymerase chain reaction.  
     
     
         12 . A method for performing nucleic acid engineering, comprising contacting single stranded DNA with a multimeric protein, wherein each unit of said multimeric protein has at least 70% sequence identity to SEQ ID NO:1, and wherein said multimeric protein binds single stranded DNA.  
     
     
         13 . A method of  claim 12 , wherein each unit of said multimeric protein has at least 80% sequence identity to SEQ ID NO:1, and wherein said multimeric protein binds single stranded DNA.  
     
     
         14 . A method of  claim 12 , wherein each unit of said multimeric protein has at least 90% sequence identity to SEQ ID NO:1, and wherein said multimeric protein binds single stranded DNA.  
     
     
         15 . A method of  claim 12 , wherein each unit of said multimeric protein has the sequence of SEQ ID NO:1, and wherein said multimeric protein binds single stranded DNA.  
     
     
         16 . A method of  claim 12 , wherein said nucleic acid engineering is selected from the group consisting of PCR-based DNA sequencing, recombination mediated cloning, PCR-mediated gene replacement, PCR-mediated recombination, RT-PCR cDNA synthesis, and in vitro sequence mutagenesis.