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US 12,258,591 B2Granted
Modified helicases
AssigneeOxford Nanopore Technologies PLC
InventorsMark Bruce, Andrew John Heron, Ruth Moysey, Szabolcs Soeroes, Elizabeth Jayne Wallace, James White
Life of this patent
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Abstract
The invention relates to a new method of characterising a target polynucleotide. The method uses a pore and a Dda helicase. The helicase controls the movement of the target polynucleotide through the pore. The invention also relates to modified Dda helicases which can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.
Claims15
A kit for characterising a target polynucleotide comprising:
- (a) a pore and
- (b) a helicase comprising:
- (i) a first polypeptide comprising the pin domain and the 1A domain from a Dda helicase and not comprising any other domains from a Dda helicase, wherein at least one cysteine residue and/or at least one non-natural amino acid have been introduced into the pin domain and/or the 1A domain, and
- (ii) a second polypeptide comprising the 2A domain, the tower domain and the hook domain from a Dda helicase and not comprising any other domains from a Dda helicase, wherein at least one cysteine residue and/or at least one non-natural amino acid have been introduced into the tower domain, wherein the helicase has the ability to control the movement of a polynucleotide;
- wherein two or more of the introduced cysteines and/or non-natural amino acids are connected to one another;
- wherein said first polypeptide is covalently attached to said second polypeptide;
- and
- wherein the pore is (1) a transmembrane protein pore, (2) a solid state pore, or (3) an oligomer pore.
5 more elements
Dependencies are inferred from the claim text.