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利用SCGE分析nZVI对DNA的损伤及其方法优化
nZVI’s DNA damage analyzed by optimized SCGE
【摘要】 本实验利用单细胞凝胶电泳技术(Single cell gel electrophoresis,SCGE)分析细胞中DNA的损伤情况,实验使用受试生物为土壤毒理学常用的模式生物赤子爱胜蚯蚓.从SCGE技术的实验方法开始,摸索SCGE的实验步骤,分别从体腔细胞的提取、铺胶、裂解、染色四个方面优化实验.探究了阴阳性的对照组,观察到阳性对照组出现条带状的DNA片段,而阴性对照组只观察到团状DNA,说明本实验的实验方法摸索成功.与此同时,将经过零价纳米铁染毒的赤子爱胜蚯蚓体腔细胞作为平行样品,进一步分析零价纳米铁(n ZVI)的生态毒理学效应,实验结果发现,在该实验条件设置下,纳米铁的急性毒性效应并不强烈,无法观察到DNA损伤.这一结论为纳米铁在环境领域的应用提供可行性的参考.
【Abstract】 Eisenia foetida,which was common used as model biology to evaluate soil toxicology,was exposed to detect its DNA damage condition by single cell gel electrophoresis(SCGE). SCGE’s procedure was explored from the following four aspects to optimize the experimental method:earthworm coelomocytes’ extraction,agarose gel’s spread coelomocytes’ disruption,and slice dyeing. Positive and negative control group were observed,clavate DNA fragments were observed in positive control group’s slice,while in negative control group only spheres DNA can be observed,which indicated that the experimental method is successfully. In the meantime,Eisenia foetida’s coelomocytes were exposed to nanoscale zero valent iron(n ZVI),and SCGE was applied to the cell,to analysis n ZVI’s ecotoxic effect. Under the experimental condition,n ZVI’s acute toxicity is small,DNA damage could not be observed. This conclusion provided feasible reference for n ZVI’s usage in the environmental field.
- 【文献出处】 天津理工大学学报 ,Journal of Tianjin University of Technology , 编辑部邮箱 ,2015年06期
- 【分类号】X171.5;Q503
- 【被引频次】1
- 【下载频次】51