Fluorescent biosensor and method for detecting transcription factor and application of fluorescent biosensor

Abstract

The invention provides a fluorescent biosensor and method for detecting a transcription factor and an application of the fluorescent biosensor and belongs to the technical field of detection analysis.The invention provides a two-way EXPAR and endonuclease IV-assisted circulatory digestion signal probe-based super-sensitive fluorescence strategy for detecting the transcription factor. Compared with conventional EXPAR, two-way EXPAR can initiate extension reaction in two directions to generate abundant trigger signals, so as to improve the amplification efficiency. The method is simple in operation, economic, high in sensitivity, low in background and good in specificity, and super-sensitive detection for the activity of the transcription factor in cells can be achieved by using the method,therefore, the method has good practical application value.

Description

technical field [0001] The invention belongs to the technical field of detection and analysis, and in particular relates to a fluorescent biosensor for detecting transcription factors and a detection method and application thereof. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Transcription factors can regulate gene expression by specifically binding to promoters / enhancers in DNA duplexes. Under the precise regulation of transcription factors, many normal biological processes, such as cell cycle progression, intracellular maintenance of metabolic and physiological balance, cell differentiation and development, and other processes can be carried out in an orderly manner....

AI Technical Summary

Problems solved by technology

[0004] In the current experiments for the detection of transcription factor activity, traditional gel electrophoresis and autoradiography detection methods mostly require complex nucleic acid labeling and gel electrophoresis procedures, which are low in sensitivity and time-consuming.

Later, new technologies for the detection of transcription factor activity were developed, including the use of dual fluorescent dye-labeled signal probes and the design of double-stem-loop molecular beacon-assisted detection, as well as various amplification strategies that have recently emerged, such as real-time polymerase chain reaction ( PCR), isothermal exponential amplification reaction (EXPAR), helicase-dependent amplification (HAD) and rolling circle amplification (RCA), etc., but these methods need special design of primers and / or templates, the inventors It was found that although these methods can detect the activity of transcription factors and screen their inhibitors to a certain extent, these methods often take a long time, require strict temperature control, and consume a large amount of high-concentration proteins and probes , the detection sensitivity still needs to be improved

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