Method for measuring cellular oxidative stress damages under ionizing radiation by using fluorescent protein as fluorescence probe

A technology of ionizing radiation and fluorescent protein, applied in the direction of fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of self-oxidation or oxidation, easy to be extinguished by light extraction, etc.

Inactive Publication Date: 2011-11-02
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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Problems solved by technology

However, there are still many shortcomings in various spectral probes for measuring intracellular ROS, such as easy to be extinguished by light, easy to be autoxidized or oxidized, non-specific localization labeling and self-participation in biological reactions to generate ROS, etc.

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  • Method for measuring cellular oxidative stress damages under ionizing radiation by using fluorescent protein as fluorescence probe
  • Method for measuring cellular oxidative stress damages under ionizing radiation by using fluorescent protein as fluorescence probe
  • Method for measuring cellular oxidative stress damages under ionizing radiation by using fluorescent protein as fluorescence probe

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Embodiment Construction

[0021] 1. Using GFP to target mitochondria to measure the early ROS generation process of cells under ionizing radiation

[0022] Mitochondria are the site of cellular oxidative phosphorylation, which provide energy for the life activities of cells by synthesizing ATP, and play a very important role in maintaining the normal physiological functions of organisms. As the only organelle with extranuclear genetic material, it has always been the focus and hotspot of radiation biology research. Mitochondria are the main source of endogenous ROS in cells, and about 1% of the oxygen in the respiratory chain generates oxygen free radicals through bypass reactions. When cells are damaged by radiation, the respiratory chain is inhibited, and the electrons leaked from the electron transport chain increase and generate more oxygen free radicals with oxygen. The increase of free radicals can aggravate the damage of various biological macromolecules in the mitochondria, reduce the function...

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Abstract

The invention discloses a method for measuring cellular oxidative stress damages under ionizing radiation by using fluorescent protein as a fluorescence probe. The content of free radicals generated in ionizing radiation is quantitatively measured by using the fluorescent protein which is selectively positioned and expressed on a subcellular structure sensitive to the ionizing radiation, so that the damages of the ionizing radiation on the specific positions of a cell are speculated by directly observing the fluorescence intensity change of the protein. By the method, the emitted fluorescence is easy to detect, has high sensitivity, stable fluorescence properties, can normally emit light in a wider pH range, has higher resistance to high temperatures, detergents, salt, organic solvents and most ordinary enzymes, is not toxic to the cell, is not interfered by false positive, makes vector construction convenient and is expressed without specificities of species, tissues and positions.

Description

technical field [0001] The invention belongs to the field of biological and biochemical detection, relates to the quantitative measurement and evaluation of cell biochemical reaction and damage caused by ionizing radiation, and proposes a fluorescent protein as an endogenous fluorescent probe for quantitatively measuring the action of free radicals produced by oxidative stress in cells in radiation reactions. Background technique [0002] Due to the rapid development of nuclear technology in the fields of industry, agriculture, and medical biology, the related nuclear safety and the impact of ionizing radiation on organisms and human health are increasingly concerned. Radiation protection, which requires accurate and quantitative detection of radiation-induced biological (cellular) processes, is still a difficult problem that is being tried to be solved. Generally, besides the direct effect of energy transfer, ionizing radiation mainly damages the body through the indirect e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 黄青柯志刚余增亮
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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