Method for controlling speed of polypeptide passing through nanopore and use thereof
AssigneeTsinghua University
InventorsJingwei Bai, Zhijie Chen
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Abstract
The present invention provides a method for controlling a speed of a polypeptide passing through a nanopore and use thereof in determining an amino acid sequence of a polypeptide. Specifically, the method comprises: conjugating a polynucleotide to the polypeptide to give a polynucleotide-polypeptide conjugate, and applying a voltage across the nanopore in the presence of a polynucleotide binding enzyme to move the conjugate through the nanopore. The polynucleotide binding enzyme controls the movement of the polynucleotide and thereby controls the movement of the conjugated polypeptide in the nanopore, thus controlling the speed of the polypeptide passing through the nanopore.
Claims20
A method for controlling a speed of a polypeptide passing through a nanopore, comprising:
- 1) conjugating the polypeptide to a polynucleotide to give a polynucleotide-polypeptide conjugate;
- and
- 2) applying a voltage across the nanopore in the presence of a polynucleotide binding enzyme to move the polynucleotide-polypeptide conjugate through the nanopore, wherein the polynucleotide binding enzyme controls the movement of the polynucleotide, thus controlling the speed of the conjugated polypeptide passing through the nanopore.
A method for measuring an electrical signal of a polypeptide formed by the polypeptide passing through a nanopore sensor, comprising:
- 1) conjugating the polypeptide to a polynucleotide to give a polynucleotide-polypeptide conjugate;
- 2) applying a voltage across the nanopore in the presence of a polynucleotide binding enzyme to move the polynucleotide-polypeptide conjugate through the nanopore, wherein the polynucleotide binding enzyme controls the movement of the polynucleotide, thus controlling the speed of the conjugated polypeptide passing through the nanopore;
- and
- 3) reading a nanopore current signal to acquire the electrical signal of the polypeptide.
A method for identifying a polypeptide or a part thereof, wherein the identification is performed by measuring an electrical signal generated by the polypeptide passing through a nanopore sensor, comprising:
- 1) ligating the polypeptide to a nucleic acid to give a nucleic acid-polypeptide ligation product;
- 2) applying a voltage across the nanopore in the presence of a nuclease to move the nucleic acid-polypeptide ligation product through the nanopore, wherein the nuclease controls the movement of the nucleic acid, thus controlling the speed of the ligated polypeptide passing through the nanopore;
- 3) reading a nanopore current signal to acquire the electrical signal of the polypeptide;
- and
- 4) identifying the polypeptide or the part thereof based on the electrical signal.
Dependencies are inferred from the claim text.