US 11,739,377 B2
Method of improving the movement of a target polynucleotide with respect to a transmembrane pore
Lakmal Nishantha Jayasinghe, Oxford (GB); Elizabeth Jayne Wallace, Oxford (GB); Jonathan Bankes Pugh, Oxford (GB); Richard George Hambley, Oxford (GB); Neil Roger Wood, Oxford (GB); Clive Gavin Brown, Cambridge (GB); James White, Oxford (GB); Andrew John Heron, Oxford (GB); Mark Bruce, Oxford (GB); Christopher Peter Youd, Oxford (GB); and Rebecca Victoria Bowen, Oxford (GB)
Assigned to Oxford Nanopore Technologies PLC, Oxford (GB)
Filed by Oxford Nanopore Technologies PLC, Oxford (GB)
Filed on Oct. 20, 2020, as Appl. No. 17/75,017.
Application 17/075,017 is a continuation of application No. 16/572,869, filed on Sep. 17, 2019, granted, now 10,844,432.
Application 16/572,869 is a continuation of application No. 15/308,252, granted, now 10,443,097, previously published as PCT/GB2015/051291, filed on May 1, 2015.
Claims priority of application No. 1407809 (GB), filed on May 2, 2014; application No. 1417708 (GB), filed on Oct. 7, 2014; and application No. 1417712 (GB), filed on Oct. 7, 2014.
Prior Publication US 2021/0139972 A1, May 13, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 14/35 (2006.01); C12Q 1/68 (2018.01); C12Q 1/6869 (2018.01)
CPC C12Q 1/6869 (2013.01) [C07K 14/35 (2013.01)] 15 Claims
 
1. A method of characterizing a target polynucleotide, comprising:
a) providing a transmembrane MspA pore and a polynucleotide binding protein in which a part of the transmembrane MspA pore which interacts with the polynucleotide binding protein and a part of the polynucleotide binding protein which interacts with the transmembrane MspA pore has been modified,
wherein the modification of the transmembrane MspA pore comprises an amino acid substitution, insertion, or deletion relative to an unmodified transmembrane MspA pore, and wherein the modification of the polynucleotide binding protein comprises an amino acid substitution, insertion, or deletion relative to an unmodified polynucleotide binding protein;
b) contacting the transmembrane MspA pore and polynucleotide binding protein provided in (a) with the target polynucleotide such that the polynucleotide binding protein controls the movement of the target polynucleotide with respect to the transmembrane MspA pore; and
c) taking one or more electrical or optical measurements as the target polynucleotide moves with respect to the transmembrane MspA pore.