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Mutant of pore protein monomer, protein pore and application thereof

A porin and mutant technology, applied in the field of characterization of target analyte characteristics, can solve the problem of single nanoporin

Pending Publication Date: 2021-11-16
QITAN TECH LTD CHENGDU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there is a single nanopore protein, and it is necessary to develop alternative nanopore proteins to realize nanopore sequencing technology

Method used

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  • Mutant of pore protein monomer, protein pore and application thereof
  • Mutant of pore protein monomer, protein pore and application thereof
  • Mutant of pore protein monomer, protein pore and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0183] In an embodiment, the wild-type porin is from Desulfovibrio thermophiles, and the amino acid sequence of the wild-type porin is SEQ ID NO:1, and the nucleotide sequence encoding the amino acid sequence is shown in SEQ ID NO:2. Mutant 1 of the porin monomer is that the wild-type porin has mutations corresponding to positions 74-81 of SEQ ID NO: 1, specifically APGNATNF at positions 74-81 is replaced by PYPANA. The protein pore of mutant 1 that includes a porin monomer is mutant pore 1. The amino acid sequence of mutant 1 of the protein monomer is shown in SEQ ID NO:24, and the nucleic acid sequence is shown in SEQ ID NO:25.

Embodiment 2

[0185] In an embodiment, the wild-type porin is from Desulfovibrio thermophiles, and the amino acid sequence of the wild-type porin is SEQ ID NO: 1, and the nucleotide sequence of the sequence encoding this amino acid is shown in SEQ ID NO: 2 . Mutant 2 of the porin monomer is that the wild-type porin has mutations corresponding to positions 74-83 of SEQ ID NO: 1, specifically APGNATNFST at positions 74-83 is replaced by AHPAATSP. The protein pore of mutant 2 that includes a porin monomer is mutant pore 2. The amino acid sequence of mutant 2 of the protein monomer is shown in SEQ ID NO:26.

Embodiment 3

[0187] In an embodiment, the wild-type porin is from Desulfovibrio thermophiles, and the amino acid sequence of the wild-type porin is SEQ ID NO: 1, and the nucleotide sequence of the sequence encoding this amino acid is shown in SEQ ID NO: 2 . Mutant 3 of the porin monomer is that the wild-type porin has mutations corresponding to positions 74-83 of SEQ ID NO: 1, specifically, APGNATNFST at positions 74-83 is replaced by PAASSSLSP. The protein pore of mutant 3 that includes a porin monomer is mutant pore 3. The amino acid sequence of mutant 3 of the protein monomer is shown in SEQ ID NO:27.

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Abstract

The invention belongs to the technical field of characterization of target analyte characteristics, and particularly provides a mutant of a pore protein monomer, a protein pore containing the mutant and application of the mutant to detection of a target analyte; wherein the amino acids of the mutant of the pore protein monomer comprise mutations at one or more positions corresponding to A74, P75, G76, N77, A78, T79, N80 and F81 of SEQ ID No: 1.

Description

technical field [0001] The invention belongs to the technical field of characterization of target analyte characteristics, and particularly relates to a mutant of a porin monomer, a protein pore containing the same and its application for detecting target analytes. Background technique [0002] With the research on the structure and sequence of nucleic acid, nucleic acid sequencing technology continues to develop, becoming the core field of life science research, and playing a huge role in promoting the development of technology in the fields of biology, chemistry, electricity, life science, and medicine. Using nanopores to develop a new type of rapid, accurate, low-cost, high-precision and high-throughput nucleic acid sequencing technology is one of the hot spots of the post-human genome project. [0003] Nanopore sequencing technology, also known as the fourth-generation sequencing technology, is a single-stranded nucleic acid molecule as a sequencing unit, using a nanopor...

Claims

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Application Information

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IPC IPC(8): C07K14/28C12N15/31C07K19/00G01N27/447G01N33/487C12Q1/6869
CPCC07K14/28C07K19/00G01N27/447G01N33/48721C12Q1/6869C12Q2565/631
Inventor 刘少伟何京雄李倩雯岳飞飞
Owner QITAN TECH LTD CHENGDU
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