Enzyme-pore constructs
AssigneeOxford Nanopore Technologies PLC
InventorsLakmal JAYASINGHE, John Hagan Pryce BAYLEY, Stephen CHELEY, Brian MCKEOWN, James WHITE, James Anthony CLARKE
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Abstract
The invention relates to constructs comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme. The pore subunit is covalently attached to the enzyme such that both the subunit and enzyme retain their activity. The constructs can be used to generate transmembrane protein pores having a nucleic acid handling enzyme attached thereto. Such pores are particularly useful for sequencing nucleic acids. The enzyme handles the nucleic acid in such a way that the pore can detect its component nucleotides by stochastic sensing.
Claims17
A construct comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme, wherein the enzyme is genetically fused to the subunit, wherein the subunit retains its ability to form a pore and is capable of forming a pore made up of 6, 7 or 8 subunits and wherein the enzyme retains its ability to handle nucleic acids.
A kit for producing a modified pore, comprising:
- (a) at least one construct comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme, wherein the enzyme is genetically fused to the subunit, wherein the subunit retains its ability to form a pore and is capable of forming a pore made up of 6, 7 or 8 subunits and wherein the enzyme retains its ability to handle nucleic acids and the remaining subunits needed to form a pore;
- or (b) at least one polynucleotide which encodes a construct comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme, wherein the enzyme is genetically fused to the subunit, wherein the subunit retains its ability to form a pore and is capable of forming a pore made up of 6, 7 or 8 subunits, wherein the enzyme retains its ability to handle nucleic acids, and wherein the polynucleotide optionally comprises the sequence shown in any one of SEQ ID NOs:
- 17, 19, 21, 23, 25, 27 and 29 or a variant thereof and polynucleotide sequences encoding any remaining subunits needed to form a pore.
Dependencies are inferred from the claim text.