Method for influencing activity of gene telomerase through serum protein bases

Abstract

The invention discloses a method for influencing the activity of gene telomerase through serum protein bases. The method comprises the following steps: step 1, splitting a to-be-detected cell, carrying out centrifuging and separating to obtain a telomerase-containing supernatant; 2, mixing the telomerase-containing supernatant with a telomerase substrate to generate a telomerase extension product;3, after the telomerase substrate extension reaction is completed, carrying out amplification reaction, and finally, forming a fluorescence detection signal; and 4, detecting the intensity of the fluorescence signal by using a luminoscope. According to the method for influencing the activity of the gene telomerase through the serum protein bases provided by the embodiment of the invention, the oligonucleotide single strand can be rapidly amplified under an isothermal condition by virtue of the extension reaction of the DNA polymerase with strand displacement activity and the property of recognizing and cutting the single strand in the specific double strand by virtue of the nucleases. The reaction not only has high amplification efficiency and can realize real-time and rapid detection, but also does not need accurate thermal cycle equipment.

Description

technical field [0001] The application relates to the field of detection of telomerase activity, in particular to a method for influencing gene telomerase activity through serum protein bases. Background technique [0002] Telomerase is a nucleoprotein reverse transcriptase composed of an RNA template and a protein catalytic subunit. Through reverse transcription, it can extend the repeat sequence (TTAGGG)n at the 3' end of the telomere of the chromosome, thereby stabilizing the length of the telomere and inhibiting cell division. In normal human tissues and cells, the activity of telomerase is inhibited, while in more than 85% of known tumor cells, telomerase often has higher activity, which is considered to be related to the activation, transformation and proliferation of tumor cells closely related. In the field of clinical diagnosis, telomerase is an important marker of malignant tumors. In addition, many cancer treatment methods and drugs also aim at inhibiting telom...