Fluorescent biosensing method for detection of interaction between small organic molecules and binding proteins based on restriction endonuclease foki inhibition assay

Abstract

The invention is a fluorescent biosensing method for detecting the interaction between organic small molecules and binding proteins based on restriction endonuclease inhibition analysis, including the interaction between the oligonucleotide DNA chain modified by the intermediate organic small molecules and the protein, restriction Conditions for the formation of endonuclease FokI dimers, selection of inhibitory sites, and real-time fluorescent quantitative detection technology based on Taqman probes. It utilizes the specific combination of small organic molecules modified in the middle of the oligonucleotide DNA chain and its binding protein or antibody, based on the inhibition of restriction endonuclease FokI digestion by the interaction between small organic molecules and proteins or antibodies, through the The cleaved Taqman probe is used for quantitative real-time fluorescence analysis to detect the interaction between small organic molecules and proteins and the small organic molecules or their binding proteins. The method has high sensitivity, simple operation and strong specificity, and can be used in the study of signal transduction and molecular regulation mechanism in biomedicine, food and agricultural product safety detection, environmental toxicant detection, drug screening, etc.

Description

technical field [0001] The invention belongs to a biosensing technology for detecting the interaction between small organic molecules and binding proteins. Interaction between nucleotide DNA chain and protein, restriction endonuclease FokI inhibition analysis, and real-time fluorescent quantitative detection method based on Taqman probe. Background technique [0002] The rapid screening and detection of organic small molecules such as organic small molecules and binding proteins, drugs and chemical toxins, and small molecule binding proteins is extremely important for clinical diagnosis, medical research, food and public safety, drug screening, environmental monitoring and other fields. meaning. At present, the commonly used detection techniques for the interaction between small organic molecules and binding proteins mainly include surface plasmon resonance, enzyme complementary fragment immunoassay, and fluorescence anisotropy analysis. These detection techniques have low...

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