US 12,258,591 B2
Modified helicases
Mark Bruce, Oxford (GB); Andrew John Heron, Oxford (GB); Ruth Moysey, Oxford (GB); Szabolcs Soeroes, Oxford (GB); Elizabeth Jayne Wallace, Oxford (GB); and James White, Oxford (GB)
Assigned to Oxford Nanopore Technologies PLC, Oxford (GB)
Filed by Oxford Nanopore Technologies PLC, Oxford (GB)
Filed on Oct. 17, 2022, as Appl. No. 18/047,019.
Application 16/893,332 is a division of application No. 15/028,651, granted, now 10,724,018, issued on Jul. 28, 2020, previously published as PCT/GB2014/052736, filed on Sep. 10, 2014.
Application 16/893,332 is a division of application No. PCT/GB2014/050175, filed on Jan. 22, 2014.
Application 18/047,019 is a continuation of application No. 16/893,332, filed on Jun. 4, 2020, granted, now 11,525,125.
Claims priority of application No. 1318464 (GB), filed on Oct. 18, 2013; application No. 1404718 (GB), filed on Mar. 17, 2014; and application No. 1406151 (GB), filed on Apr. 4, 2014.
Prior Publication US 2023/0212535 A1, Jul. 6, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/14 (2006.01); C12N 9/90 (2006.01); C12Q 1/6869 (2018.01)
CPC C12N 9/14 (2013.01) [C12N 9/90 (2013.01); C12Q 1/6869 (2013.01); C12Q 2565/631 (2013.01); C12Y 306/04012 (2013.01)] 15 Claims
 
1. 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.