环境与职业医学2017，Vol.34Issue(6)：511-516,6.DOI:10.13213/j.cnki.jeom.2017.17210
荧光转基因斑马鱼Tg(ere-vtg:egfp)质粒的构建及验证
Generation and verification of Tg(ere-vtg:egfp) plasmid in fluorescent transgenic zebrafish
摘要
Abstract
[Objective] To construct a fluorescent transgenic zebrafish model sensitive to 17α-ethynylestradiol (EE2) to directly monitor environmental estrogens (EEs) pollution in aquatic system.[Methods] Tg(ere-vtg:egfp) recombined plasmid was constructed,linearized and microinjected into 1-2 cell zebrafish embryos.The injected embryos were exposed to 10ng/L EE2 to induce green fluorescent protein (GFP),and further raised till sexually mature to build transgenic zebrafish line.The sensitivity was tested with protocols using homozygote larvae and embryos exposed to different concentrations of EE2.The specificity was tested using several environmental chemicals with similar activity or structure.[Results] Tg(ere-vtg:egfp) transgenic zebrafishes was successfully generated after microinjection with Tg(ere-vtg:egfp) plasmid and genetic screening.GFP expression in Tg(ere-vtg:egfp) transgenic zebrafish line was related to the concentration and time of EE2 exposure.GFP expression was induced at 0.1 ng/L EE2 (131 hpf) in zebrafish embryos.Tg(ere-vtg:egfp) transgenic zebrafish larvae expressed GFP after exposure to estrogens with varied structures but not after exposure to non-estrogen chemicals.[Conclusion] Using Tg(ere-vtg:egfp) transgenic zebrafish can monitor EEs pollution dynamically.关键词
环境雌激素/转基因斑马鱼/卵黄蛋白原/绿色荧光蛋白/环境监测Key words
environmental estrogen/transgenic zebrafish/vitellogenin/green fluorescent protein/environment monitoring分类
医药卫生引用本文复制引用
孙冰,许慧慧,夏敏杰,丁训城,胡晶莹,李卫华..荧光转基因斑马鱼Tg(ere-vtg:egfp)质粒的构建及验证[J].环境与职业医学,2017,34(6):511-516,6.基金项目
国家自然科学基金项目(编号:31540038) （编号:31540038）
上海市科委实验动物研究专项(编号:15140901200) （编号:15140901200）
上海市第四轮公共卫生三年行动计划重点学科建设计划—环境卫生学与劳动卫生学(编号:15GWZK0201) （编号:15GWZK0201）
上海市青年科技英才扬帆计划(编号:15YF1410000) （编号:15YF1410000）
