Polymerase compositions and methods
Disclosed herein are modified polymerase compositions exhibiting altered polymerase activity, which can be useful in a variety of biological applications. Also disclosed herein are methods of making and using such compositions. In some embodiments, the compositions exhibit altered properties that can enhance their utility in a variety of biological applications. Such altered properties, can include, for example, altered nucleotide binding affinities, altered nucleotide incorporation kinetics, altered photostability and/or altered nanoparticle tolerance, as well as a range of other properties as disclosed herein.
1. A mutant DNA polymerase comprising the amino acid sequence of SEQ ID NO:7.
2. The mutant DNA polymerase of claim 1 , which exhibits reduced 3′ to 5′ exonuclease activity compared to a wild type DNA polymerase which comprises the amino acid sequence of SEQ ID NO:6.
3. The mutant DNA polymerase of claim 1 , further comprising a peptide tag which includes 2-8 consecutive histidine residues, wherein the peptide tag is linked to the N-terminal or C-terminal end of the mutant DNA polymerase.
4. The mutant DNA polymerase of claim 3 , wherein the peptide tag further comprises zinc, nickel, cobalt or copper ions.
5. The mutant DNA polymerase of claim 1 , further comprising a peptide tag which includes the amino acid sequence MNHLVHHHHHHIEGRHMELGTLEGS (SEQ ID NO:51), wherein the peptide tag is linked to the N-terminal or C-terminal end of the mutant DNA polymerase.
6. The mutant DNA polymerase of claim 1 , further comprising a peptide tag which includes the amino acid sequence MHHHHHHKH (SEQ ID NO:52), wherein the peptide tag is linked to the N-terminal or C-terminal end of the mutant DNA polymerase.
7. The mutant DNA polymerase of claim 1 , further comprising a peptide tag which includes the amino acid sequence LLGAAAKGAAAKGSAA (SEQ ID NO:16), wherein the peptide tag is linked to the N-terminal or C-terminal end of the mutant DNA polymerase.
8. The mutant DNA polymerase of claim 1 , further comprising a peptide tag which includes the amino acid sequence LLGGGGSGGGGSAAAGSAA (SEQ ID NO:17), wherein the peptide tag is linked to the N-terminal or C-terminal end of the mutant DNA polymerase.
9. The mutant DNA polymerase of claim 1 , further comprising a peptide tag which includes the amino acid sequence GLNDIFEAQKIEWHE (SEQ ID NO:53), wherein the peptide tag is linked to the N-terminal or C-terminal end of the mutant DNA polymerase.