MUTANT DNA POLYMERASES
Provided herein are mutant DNA-dependent polymerases which are derived from, or otherwise related to, wild type RB69 DNA polymerase. These mutant polymerases are capable of selectively binding labeled nucleotides. These mutant polymerases are also capable of incorporating a variety of naturally occurring and modified nucleotides, including, for example, terminator nucleotides.
1 . An isolated mutant DNA polymerase capable of selectively transiently-binding a labeled nucleotide and selectively incorporating a terminator nucleotide.
2 . The isolated mutant DNA polymerase of claim 1 , comprising a mutant RB69 DNA polymerase.
3 . The isolated mutant DNA polymerase of claim 1 , comprising an amino acid sequence according to any one of SEQ ID NOS:2-8.
4 . The isolated mutant DNA polymerase of claim 1 further comprises a reporter moiety.
5 . The isolated mutant DNA polymerase of claim 4 , wherein the reporter moiety is an energy transfer donor moiety.
6 . The isolated mutant DNA polymerase of claim 5 , wherein the energy transfer donor moiety is a fluorescent dye or a nanoparticle.
7 . A system comprising a mutant DNA polymerase of claim 1 bound to a DNA template and a primer, and a nucleotide transiently-bound to the mutant DNA polymerase.
8 . A nucleic acid molecule encoding the mutant DNA polymerase of claim 1 .
9 . The nucleic acid molecule of claim 8 which is DNA or RNA.
10 . A vector comprising the nucleic acid molecule of claim 8 .
11 . The vector of claim 10 further comprising a promoter sequence joined with the nucleic acid molecule encoding the mutant DNA polymerase.
12 . The vector of claim 11 , wherein the promoter is constitutive or inducible.
13 . A host cell carrying the vector of claim 11 .
14 . The host cell of claim 13 which is a phage, a prokaryote cell or a eukaryote cell.
15 . A method for producing a mutant DNA polymerase polypeptide, comprising culturing the host cell of claim 14 under conditions suitable for the host cell to produce the mutant DNA polymerase polypeptide.
16 . The mutant DNA polymerase produced by the method of claim 15 .
17 . An isolated mutant DNA polymerase comprising an amino acid sequence that is at least 80%, 85%, 90%, 95%, 97% or 99% identical to any amino acid sequence selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7 and SEQ ID NO:8.