The Preliminary Study On The Bioassay Of Dioxin-like Compounds

Dioxin-like compounds(DLCs)are typical persistent organic pollutants that have a health threat to the organism with strong carcinogenic,endocrine disrupting,hepatotoxicity,immunotoxicity,skin toxicity,and fertility disorders,so monitoring them is extremely important.At present,there are many analytical methods of dioxin like compounds,but they can be summed up as instrumental analysis and biological analysis.Among them,the sample pretreatment process of instrument analysis is complicated,the detection cycle is long,the detection cost is high,and the technical requirements of the testing personnel are relatively high,which limits the application of the method in environmental monitoring.Because of the similar toxic mechanism of dioxin-like compounds,them can influence the expression of cyplal gene through AhR signaling pathway.Therefore,we designed and constructed the cyplal promoter to regulate GFP gene,and obtained stably transfected hepG2 cells line and transgenic zebrafish,Try to use in vitro and in vivo bioanalysis of dioxin-like compounds for detection and analysis.Preliminary results are as follows:1)In cultured cells in vitro,the modified cyplal promoter,which contains multiple dioxin response elements(DRE),can indeed increase the expression of TCDD-induced reporter genes;2)Cell lines constructed with GFP as a reporter gene,because of their high background fluorescence intensity,can show differences in fluorescence intensity only when treated with far higher concentrations than those in the environment,so the cell line is not suitable for Detection of dioxin in the environmert;3)Transgenic zebrafish with GFP gene regulated by the modified cyplal promoter was successfully obtained by TALEN.The results of preliminary detection of TCDD using the transgenic zebrafish showed:The fluorescence intensity of the transgenic zebrafish showed a good dose effect and time effect under TCDD exposure,and the sensitivity of the test is high,which provides an important basis for the further screening of transgenic fish for further monitoring the level of dioxin-like compounds in the water environment.