Fluorescent biosensing method for analyzing and detecting mutual action of organic micromolecules and combined protein based on T7 exonuclease inhibition

Abstract

The invention provides a fluorescent biosensing method for analyzing and detecting mutual action of organic micromolecules and combined protein based on a T7 exonuclease inhibition. The method comprises mutual action of an oligonucleotide DNA (Deoxyribonucleic Acid) single chain with the middle part modified by organic micromolecules and protein, selection of a T7 exonuclease inhibition site, and a real-time fluorescent quantitative detection technique based on a Taqman probe. The method provided by the invention can be used for detecting the mutual action of the organic micromolecules and the protein, and the organic micromolecules or conjugated protein thereof through the fluorescent real-time quantitative analysis of the Taqman probe subjected to enzyme digestion based on the inhibition effect of the mutual action of the organic micromolecules and the protein or an antibody to T7 exonuclease by utilizing the specific binding of the organic micromolecules modifying the middle part of the oligonucleotide DNA chain and the conjugated protein or antibody thereof. The method provided by the invention is high in sensitivity, simple and convenient in operation and strong in specificity, and can be used for signal transduction and molecule regulation and control mechanism research in biomedicine, food and farm product security detection, environmental poison detection,drug screening and the like.

Description

technical field [0001] The invention belongs to a biosensing technology for detecting the interaction between small organic molecules and binding proteins, which specifically refers to the interaction between oligonucleotide DNA chains and proteins modified in the middle of small organic molecules, the selection of inhibitory sites, T7 nucleic acid Exonuclease inhibition analysis, and real-time fluorescence quantitative detection method based on Taqman probe. Background technique [0002] The rapid screening and detection of small organic molecules such as interactions between small organic molecules and binding proteins, drugs and chemical toxins, and small organic molecule binding proteins are extremely useful for clinical diagnosis, medical research, food and public safety, drug screening, and environmental monitoring. Significance. At present, the commonly used detection techniques for the interaction between small molecules and binding proteins mainly include surface ...

AI Technical Summary

Problems solved by technology

These detection techniques have low sensitivity, poor reliability, and require sophisticated instruments, which cannot meet the needs of accurate and rapid detection