GFP遗传改造四膜虫细胞的构建及其对重金属污染的荧光响应.docVIP

GFP遗传改造四膜虫细胞的构建及其对重金属污染的荧光响应 梁海霞 刘红伟 太原理工大学信息工程学院 太原理工大学化学化工学院 X 关注成功！ 加关注后您将方便地在 我的关注中得到本文献的被引频次变化的通知！ 新浪微博 腾讯微博 人人网 开心网 豆瓣网 网易微博 摘????要： 为了获得可用于检测环境中多种重金属污染的遗传改造生物细胞, 将具有重金属诱导特性的金属硫蛋白基因MTT1启动子和外源报告基因绿色荧光蛋白gfp的融合序列, 通过基因枪法转染嗜热四膜虫细胞。经同源重组和巴龙霉素抗性筛选, 获得稳定的GFP遗传改造四膜虫细胞株。该细胞株经Cd2+诱导, MTT1启动子可启动外源基因gfp正确表达, 在蓝光激发下可发绿色荧光。GFP遗传改造四膜虫细胞对重金属离子的荧光响应效应由大到小依次为Cd2+, Cr6+, Hg2+;响应浓度范围和具有剂量-效应关系的浓度范围由大到小依次为Cd2+, Hg2+, Cr6+;最低检测限由大到小依次为Cr6+, Hg2+, Cd2+, 而对Cu2+无荧光响应效应。因此, GFP遗传改造四膜虫可作为环境中Cd2+、Hg2+和Cr6+污染的生物检测细胞。 关键词： 金属硫蛋白; 绿色荧光蛋白; 遗传改造四膜虫; 重金属污染; 荧光响应; 作者简介：梁海霞 (1983-) , 女, 博士, 讲师, 主要从事生物工程技术研究, (E-mail) lianghaixia@tyut.edu.cn 收稿日期：2017-09-12 基金：国家自然科学青年基金资助项目 Construction of Genetically Modified Tetrahymena with GFP and Its Fluorescence in Response to Environmental Heavy Metal Pollution LIANG Haixia LIU Hongwei College of Information Engineering, Taiyuan University of Technology; College of Chemistry and Chemical Engineering, Taiyuan University of Technology; Abstract： To obtain genetically modified cells which can be used to detect a variety of environmental heavy metals pollution, the fusion gene containing heavy metals induced metallothionein gene MTT1 promoter and exogenous gfp reporter gene was transfected into Tetrahymena thermophilia by biolistic bombardment.By homologous recombination under increasing paromomycin, the genetically modified Tetrahymena with GFP was constructed.With treatment of Cd2+, the MTT1 promoter can initiate the correct expression of exogenous gfp gene of the genetically modified Tetrahymenain which green fluorescence can be induced by blue light excitation.The fluorescence response of the genetically modified Tetrahymena with GFP to heavy metals was in order of Cd2+, Cr6+and Hg2+.And the concentration ranges within which fluorescence was induced and with dose-response relationship were both in order of Cd2+, Hg2+and Cr6+.The detection limit of heavy metals was in order of Cr6+, Hg2+and Cd2+.However, the genetically modified