Method for detecting protein or protein complex by identifying a plurality of epitopes synchronously

Abstract

The invention adopts oligonucleotide aptamers for identifying special tripeptide sequence KAI, GEL, DGI and LAS, extanding oligonucleotide aptamers by Structure-switch and phosphorylation decorating 5' end thereof, synthetising four connexons; connecting oligonucleotide aptamers combinated with target albumen through proximity dependent DNA Ligation technique as chain ringlike DNA; amplifying detecting signals via rolling-circle DNA duplicate method; the technique can detect target albumen sensitively, the least detecting concentration can get nM level; this method with flexibility can adjust according needed numbers of ligand; can detect albumen composite, provide sensitive detecting method for chip technique using oligonucleotide aptamers as ligand.

Description

technical field [0001] The invention relates to proteomics research technology, in particular to a method in which an oligonucleotide aptamer (Nucleic Acid Aptamer) that recognizes protein epitopes is used to simultaneously recognize multiple epitopes to detect proteins or protein complexes. Background technique [0002] Protein expression can exist as a monomer or a dimer, and there are more complex complexes formed by multiple proteins. The detection of these proteins and even complexes is very important, especially in proteomics research , we not only need to know what kind of protein is expressed, but we also need to know the state of the protein, including modifications and complexes. Therefore, the detection of proteins, especially the detection of various states of proteins after translation is very important. The study of proteomics requires high-throughput technology and sensitive detection methods. Recently, two-dimensional electrophoresis technology and protein c...

AI Technical Summary

Problems solved by technology

[0003] In protein detection, the method of proximity dependent DNA ligation can detect trace amounts of protein, but because the final quantitative method it adopts is the method of fluorescent quantitative PCR, it is often caused by the non-specificity of PCR amplification or the adaptation of oligonucleotides. The GC content of the daughter is too high to make the PCR reaction impossible, so the auxiliary fragment has to be extended

In addition, this method can only detect the same epitope in binary complexes of the same protein

Of course, the main disadvantage of this method is the presence of background ligation reactions, which require tedious optimization of reaction conditions to minimize interference

This method can only detect two epitopes at most at the same time, so more flexible methods are needed for proteomics research

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