Reporter plasmid for biological detection of radioactive ray dose and construction method thereof

A reporting plasmid and biological detection technology, applied in biochemical equipment and methods, microbial measurement/inspection, dosimeter, etc., can solve the problems of complicated operation, difficult to quantify accurately, high cost of detection, etc., and achieve the goal of improving accuracy Effect

Inactive Publication Date: 2009-01-21
ZHEJIANG UNIV
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AI Technical Summary

Problems solved by technology

[0003] At present, there are many methods for biological detection of radiation dose. Commonly used are: the relationship between the comet tail formed by DNA damage after radiation exposure and the comet core expressed by undamaged cells to study single-cell gel electrophoresis for predicting radiation dose (Comet test) method (Berwick and Vineins, 2000); there is also a chromosomal aberration analysis method (Ballarini and Ottolenghi, 2003; Duran et al., 2002; Camparoto et al., 2003), but the common disadvantages of these methods are complex operation, high detection cost, more human factors in damage observation, and difficult to quantify accurately, etc. Disadvantages: Although the use of auxiliary means such as computers can properly improve the accuracy of its measurement, these methods can only detect and measure low-dose radiation, and there is no method for biological detection and measurement of high-dose radiation

Method used

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Embodiment Construction

[0020] The present invention will be described in detail below based on the examples, but the present invention is not limited to these examples.

[0021] (1) Construction of shuttle vector plasmid

[0022] Plasmid pUE30 was extracted from radiation-resistant bacteria (Deinococcus radiopugnance ATCC19172) and digested with SphI; then digested with SphI to contain the replication initiation domain of E. coli vector pUC19, the active Amp resistance gene in E. Plasmid pKatCAT (Funayama et al., Mutat. Res., 435:151-161, 1999) with an active chloramphenicol resistance gene in it was mixed with the above SphI digest of pUE30 and ligated with DNA ligase. The above linker was transformed into Escherichia coli JM109 strain by electroporation. A strain having the pUE30 and pKatCAT-linked plasmids was selected from the Amp-resistant transformants to obtain a shuttle vector plasmid.

[0023] (2) Construction of reporter plasmid

[0024] Using the genomic DNA of the radiation-resistant ...

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Abstract

The invention relates to a reporter plasmid in radiation dose used for biological detection and a preparation method thereof. The preparation method comprises the following steps: a shuttle vector between radiation resistant bacteria and coliform bacteria is taken as a foundation so as to be recombined with a DNA fragment in the luciferase gene lux field derived from the host DNA of a North American fire fly, thereby forming a reporter plasmid of the shuttle vector which has the activity of luciferase genes and is positioned between the radiation resistant bacteria and the coliform bacteria; the reporter plasmid is fed into a wild type R1 for transforming the radiation resistant bacteria so as to obtain transformant radiation resistant bacteria which are irradiated by gamma ray; the transformant radiation resistant bacteria are cultured to reach a stationary phase, and are irradiated by gamma rays so as to be cultured again; and according to the luminous quantity of a character transformant, different luminous quantities irradiated by the gamma rays in different doses are measured. Research result shows that within the adopted dose range of between 0 and 2, 000Gy, the correlation coefficient between the luminous quantities of thalli irradiated by the gamma rays in different luminous quantities and irradiation doses is equal to 0.892, reaching the extremely significant level.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a gene plasmid and a construction method thereof, that is, a reporter plasmid for biological detection of radiation dose and a construction method thereof. Background technique [0002] The health hazards of ionizing radiation and radiation safety assessment, the classification, diagnosis and treatment of acute and chronic radiation injuries under various conditions all need to understand and determine the exposure dose. The biological detection of radiation dose is the use of biological materials such as tissues, cells, DNA, proteins, etc. after ionizing radiation, which has a certain amount-effect relationship with the radiation dose. A class of biomarkers and analytical methods used to measure radiation doses through biological changes and analysis. The greatest advantage of using living organisms for dosimetry compared to physical detection is directness and faithful representatio...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12Q1/66C12N15/63G01T1/02
Inventor 屠振力
Owner ZHEJIANG UNIV
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