Protein/polypeptide sequencing method adopting Aerolysin nanopores

A nano-pore, protein technology, applied in the biological field, can solve the problems of inability to effectively identify 20 amino acids, difficulty in obtaining amino acid sequence information, lack of efficient organic fluorophores, etc., and achieve the effect of improving amino acid identification ability.
CN112480204APending Publication Date: 2021-03-12NANJING UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NANJING UNIV
Publication Date
2021-03-12

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Abstract

The invention provides a method for protein / polypeptide sequencing based on Aerolysin nanopores. The method realizes specific resolution of natural amino acid and post-translational modification thereof and accurate acquisition of a single-molecule protein sequence, and comprises the following steps: (1) carrying out protein unfolding; (2) marking an end position sequencing starting point; (3) carrying out electrified preliminary screening; (4) unfolding a polypeptide tertiary structure; (5) carrying out amino acid orthogonal recognition; (6) carrying out limited range perturbation assisted amino acid recognition; and (7) carrying out single molecule protein sequence determination. According to the method, an innovative method for accurate measurement of single protein molecule amino acidsequence and post-translational modification is established for sensitive detection of information of 20 amino acid sequences.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a protein / polypeptide sequencing method based on Aerolysin nanopores. Background technique

[0002] Thousands of different proteins maintain all the functions of cells. The precise determination of the amino acid sequences of proteins in organisms can provide a basis for understanding protein functions, the biological processes they participate in, and the interactions between proteins and proteins (or other biomolecules). Key Information. According to the transmission direction of genetic information, protein synthesis is synthesized by processes such as DNA transcription, post-transcriptional processing, translation, and post-translational modification. However, a gene can be spliced ​​in multiple mRNA forms during transcription, and the same protein can undergo post-translational modification in many forms, so there is a large difference between the genotype and pheno...

Claims

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