Medical Research Council

United Kingdom Research and Innovation

Philipp Holliger

Vitor B Pinheiro

Farid Ghadessy

Christopher Cozens

Alexander I Taylor

Chris Cozens

Marc d'Abbadie

Nicola Ramsay

Ann-Sofie Jemth

Zoryana Oliynyk

Jennifer Lee Ong

Marc D'Abbadie

Claudia Baar

Marc D'abbadie

The invention relates to a nucleic acid polymerase capable of producing a non-DNA nucleotide polymer from a DNA nucleotide template, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at one or more residues of the thumb region, said residues selected from: amino acids 651 to 679 (patch 10A); wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue E664. In one embodiment said polymerase comprises the mutations Y409G and E664K. In one embodiment said polymerase comprises amino acid sequence corresponding to SEQ ID NO:12. The invention also relates to A nucleic acid polymerase capable of reverse transcribing a HNA nucleotide polymer into a DNA nucleotide polymer, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of sEQ ID NO:1, wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue I521.

Enzymes

The invention relates to a nucleic acid polymerase capable of producing a non-DNA nucleotide polymer from a DNA nucleotide polymer template, the polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO: 1, wherein the amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO: 1, wherein the amino acid sequence comprises the mutations P657T, E658Q, K659H, Y663H, D669A, K671N, and T676I; wherein the amino acid sequence is further mutated relative to the amino acid sequence of SEQ ID NO: 1 at residue: E664 and wherein the amino acid sequence comprises the mutation E664K. The invention also relates to methods of making nucleotide polymers comprising use of this polymerase. Suitably the nucleotides are arabino nucleotides such as ARA or FANA nucleotides.

Polymerase capable of producing non-DNA nucleotide polymers

An engineered DNA polymerase characterised in that the polymerase exhibits an enhanced ability to process nucleic acid in the presence of environmental and biological inhibitors compared to wild type DNA polymerase.

Polymerase

The invention relates to a nucleic acid polymerase capable of producing a non-DNA nucleotide polymer from a DNA nucleotide polymer template, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at one or more residues of the thumb region, said residues selected from: amino acids 651 to 679 (patch 10A); wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue E664. In one embodiment said polymerase comprises the mutations Y409G and E664K. In one embodiment said polymerase comprises amino acid sequence corresponding to SEQ ID NO:12. The invention also relates to A nucleic acid polymerase capable of reverse transcribing a HNA nucleotide polymer into a DNA nucleotide polymer, said polymerase comprising amino acid sequence having at least 36% identity to the amino acid sequence of SEQ ID NO:1, wherein said amino acid sequence is mutated relative to the amino acid sequence of SEQ ID NO:1 at residue I521.

The present invention relates to DNA polymerases. In particular the invention relates to a method for the generation of DNA polymerases exhibiting a relaxed substrate specificity. Uses of mutant polymerases produced using the methods of the invention are also described.

DNA polymerase

An emulsion is useful in allowing a wide variety of gene products to be expressed via eukaryotic in vitro expression. The emulsion comprises a silicone based surfactant, a hydrophobic phase and a hydrophilic phase; wherein the hydrophilic phase comprises a plurality of compartments containing a functional in vitro eukaryotic expression system.

Emulsion compositions

The present invention relates to the development of a novel method for the selection of nucleic acid processing and other enzymes. In particular the invention relates to a method for the selection of nucleic acid polymerases and other enzymes with desired properties based on the method of compartmentalized self-tagging.

Growing community of inventors

Cambridge, United Kingdom

Philipp Holliger