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A radiation biological dosimeter, preparation method and application

A technology of radiation biological dose and Escherichia coli, which is applied in the fields of radiation biological dosimeter, preparation and application, can solve the problems of long detection time, high detection background, and many interference signals, achieve stable and bright fluorescence, simplify operating procedures, and improve Effect of Detection Sensitivity

Active Publication Date: 2021-09-24
BEIJING INSTITUTE OF TECHNOLOGYGY
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Problems solved by technology

[0013] In view of the above-mentioned existing technologies only for the detection of chemical toxic reagents, less attention is paid to radiation factors, and the detection time of the Ames method in the previous detection methods is long, and the chemical sensor detection method has many interference signals, and there are high detection backgrounds and high sensitivity. Low defect, one of the purposes of the present invention is to provide a radiation biological dosimeter, which uses genetically engineered bacteria as a microbial sensor, and directly reflects the radiation dose through the fluorescent signal, and effectively uses the group effect to amplify the signal, innovatively established A radiation biological dosimeter. Compared with other chemical dosimeters, the radiation biological dosimeter has many advantages: non-toxic and harmless, convenient for online detection and preparation, low cost, and the interaction between organisms can be used. The group effect among microorganisms can realize signal amplification, facilitate detection, and improve the detection sensitivity of radiation biological dosimeters

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  • A radiation biological dosimeter, preparation method and application
  • A radiation biological dosimeter, preparation method and application
  • A radiation biological dosimeter, preparation method and application

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Embodiment 1

[0069] A preparation method of the radiation biological dosimeter according to the present invention, the steps of the preparation method are as follows:

[0070] (1) Using the Escherichia coli MG1655 genome as a template, the SoxR promoter of the target gene was amplified by PCR; the Cda promoter of the target gene was amplified by PCR using the genome of the plasmid pColD-CA23 as a template; GENEWIZ) synthesized the pUC 19 cloning plasmid (Escherichia coli, Cloning vector pUC19) genome containing the EGFP gene sequence optimized by codon degeneracy as a template, and the target gene EGFP gene was amplified by PCR; the PCR amplification product was subjected to agarose For gel electrophoresis separation, the Marker is Marker III. After the separation, the gel is cut and recovered to obtain the DNA of the target gene fragment.

[0071] The nucleotide sequence of the PUC 19 clone plasmid genome is SEQ ID No.3 in the nucleotide sequence list, specifically as follows:

[0072] 5...

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Abstract

The invention relates to a radiation biological dosimeter, a preparation method and an application, and belongs to the field of biotechnology. The biological dosimeter is a SoxR radiation biological dosimeter formed by adding the EGFP gene downstream of the SoxR promoter of Escherichia coli or a Cda radiation biological dosimeter formed by adding the EGFP gene downstream of the Cda promoter of Escherichia coli. The radiation biological dosimeter can monitor the physical radiation dose in real time online, and has a wide range of detection and application; the group effect of microorganisms is used as a biological signal amplification system, and the sensitivity is greatly improved compared with the existing related detection technology; The above-mentioned radiation biological dosimeter is placed in the microfluidic chip for cultivation, which can realize online real-time monitoring, realize commercial production, is easy to carry, and can detect quickly. for reference.

Description

technical field [0001] The present invention relates to a kind of radiation biological dosimeter, preparation method and application, in particular, to a kind of biometric method using biotechnology to modify the gene sequence of microorganisms as the radiation dose, for the low linear energy transfer (LinearEenergy Transfer, LET) physical Radiation and ultraviolet radiation are monitored, the detection time is short, the results are intuitive and visible, and there is a certain dose-effect relationship between the response intensity and the radiation dose. The fluorescence intensity value reflects the cytotoxicity of the radiation in the environment, and it is harmful to chemical cells. The reagent has response and detection broad-spectrum; it belongs to the field of biotechnology. Background technique [0002] Water pollution, noise pollution, air pollution and radiation are known as the four major environmental pollution. According to the biological effects of radiation,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/65C12N1/21C12N15/113G01N21/64C12R1/19
CPCC07K14/245C12N15/65G01N21/6486
Inventor 张永谦郝晓婷邓玉林
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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