Modified helicases
AssigneeOxford Nanopore Technologies PLC
InventorsAndrew John Heron, James Anthony Clarke, Ruth Moysey, Elizabeth Jayne Wallace, Mark John Bruce, Lakmal Jayasinghe, Domenico Caprotti, Szabolcs Soeroes, Luke McNeill, David Antoni Alves, Rebecca Victoria Bowen, John Milton
Life of this patent
Estimating the term…
Abstract
The invention relates to modified helicases with reduced unbinding from polynucleotides. The helicases can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.
Claims7
A helicase formed from one or more monomers and comprising a polynucleotide binding domain which comprises in at least one conformational state an opening through which a polynucleotide can unbind from the helicase, wherein the helicase is modified such that two or more parts on the same monomer of the helicase are connected to reduce the size of the opening and wherein the helicase retains its ability to control the movement of the polynucleotide.
2-22. (canceled)
A Hel308 helicase in which one or more cysteine residues and/or one or more non-natural amino acids have been introduced at one or more of the positions which correspond to D272, N273, D274, G281, E284, E285, E287, S288, T289, G290, E291, D293, T294, N300, R303, K304, N314, S315, N316, H317, R318, K319, L320, E322, R326, N328, S615, K717, Y720, N721 and S724 in Hel308 Mbu (SEQ ID NO:
- 10), wherein the helicase retains its ability to control the movement of a polynucleotide.
24-49. (cancled)
A kit for characterising a target polynucleotide comprising (a) a pore and (b) a helicase formed from one or more monomers and comprising a polynucleotide binding domain which comprises in at least one conformational state an opening through which a polynucleotide can unbind from the helicase, wherein the helicase is modified such that two or more parts on the same monomer of the helicase are connected to reduce the size of the opening and wherein the helicase retains its ability to control the movement of the polynucleotide.
52-59. (canceled)
Dependencies are inferred from the claim text.