Mutant of pore protein monomer, protein pore, application of mutant and application of protein pore

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: 2022-01-11
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, application of mutant and application of protein pore
  • Mutant of pore protein monomer, protein pore, application of mutant and application of protein pore
  • Mutant of pore protein monomer, protein pore, application of mutant and application of protein pore

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Experimental program
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Effect test

Embodiment 1

[0188] In an embodiment, the wild-type porin is from Microvesicle thermostable, 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 multiple mutations corresponding to SEQ ID NO: 1, specifically Y76F, S79A, S83T, and F84N. 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:16.

Embodiment 2

[0190] In an embodiment, the wild-type porin is from Microvesicle thermostable, 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 multiple mutations corresponding to SEQ ID NO: 1, specifically R50K, Y76F, K77R, S79Y, S83A, and F84G. 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:17.

Embodiment 3

[0192] In an embodiment, the wild-type porin is from Microvesicle thermostable, 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 multiple mutations corresponding to SEQ ID NO: 1, specifically R50K, Y76F, K77R, S79L, S83A, and F84Q. 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:18.

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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 an application of the mutant or the protein pore to detection of a target analyte. The amino acid of the mutant of the pore protein monomer comprises a sequence as shown in SEQ ID NO: 1 or a sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 80%, 70%, 60% or 50% identity with the sequence as shown in SEQ ID NO: 1, and the amino acid of the mutant of the pore protein monomer comprises mutations at one or more positions corresponding to Y76, S79, S83 and F84 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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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/195C12N15/31C07K19/00G01N27/447G01N33/487C12Q1/6869
CPCC07K14/195C07K19/00G01N27/447G01N33/48721C12Q1/6869C12Q2565/631
Inventor 刘少伟谢馥励李倩雯赵帅
Owner QITAN TECH LTD CHENGDU
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