Development method of fluorescent protein transgenic zebra fish capable of highly sensitively monitoring water body PAHs by utilizing cascade amplification effect

Abstract

The invention belongs to the field of biotechnology, and in particular relates to a development method of fluorescent protein transgenic zebra fish capable of highly sensitively monitoring water body PAHs by utilizing the cascade amplification effect. According to the method, a UAS-Gal4 gene expression regulation system is taken as a basis, a multiring aromatic hydrocarbon response element AhREs after optimization and reorganization is utilized for driving the expression of the upstream plasmid Ga14, and further, the expression of the fluorescent gene eGFP in the downstream UAS plasmid is regulated; meanwhile, the TALE-TF transcriptional activation enhancement technology is innovatively combined, the expression quantity of the eGFP gene is further enhanced by utilizing the TALE transcriptional activation effect, and then the purpose of increasing the fluorescence signals through multiple ways is achieved; finally, the low-copy induced fluorescent protein transgenic zebra fish with stable inheritance is obtained by utilizing the I-SceI wide-range nuclease transfer system, and the influences on the toxicity of the zebra fish are small; by manufacturing the fluorescent protein transgenic zebra fish highly sensitive to the environmental pollutants, the toxic pollutants in the water body can be easily, efficiently, sensitively and intuitively monitored.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for developing fluorescent protein gene-transferred zebrafish by using cascade amplification effect to monitor water body PAHs pollutants with high sensitivity. Background technique [0002] At this stage, environmental pollution, especially water pollution, is widespread all over the world. Water pollution will not only affect the survival and reproduction of aquatic organisms, but also pollutants will reach the end of the food chain through bioaccumulation and concentration, seriously affecting human health. In recent years, people have frequently detected polycyclic aromatic hydrocarbons and other organic pollutants in the environment. Studies have shown that such pollutants have caused serious threats to the survival of organisms in the water environment, soil and sediments. Among them, the organic pollutants in the environmental water body are mainl...

AI Technical Summary

Problems solved by technology

In 2001, Mattingly et al first confirmed that zebrafish transcription factors can recognize human AhREs and can drive the expression of GFP under the induction of TCDD, but they only studied the functions of aryl hydrocarbon receptors and their downstream genes, and did not apply them to In vivo monitoring of aromatic hydrocarbon pollutants

Since then, Nebert et al. used the promoter containing AhREs to drive the expression of LUC reporter gene, and developed transgenic zebrafish to detect aromatic hydrocarbon organic pollutants in water, but the LUC reporter gene in the transgenic zebrafish they developed could not be transmitted to F 2 generation

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