基因重组细胞监测重金属污染物的实验研究.pdfVIP

中国环境科学 2007，27(6)：835～840 China Environmental Science 基因重组细胞监测重金属污染物的实验研究 李湘呜 ，罗方妮 ，陈春波。，吴玉娟 ，李华玲 (1．扬州大学医学院，江苏扬州225001；2．扬州大学动物科学 与技术学院，江苏 扬州225001；3．扬州大学兽医学院，江苏 扬州 225001) 摘要：研究用基因重组细胞快速评价重金属污染物对健康的危害，采用分子生物学方法，将亲电性化学反应元件(EPRE)与TK基本启动子和 编码虫荧光素酶(LUC)基因相融合，构建成EPRE调控的LUC报告载体，将其转染于HeLa细胞．并将该细胞作用于不同浓度的NaAsO2、 CdC12和HgC12，用荧光素酶试剂盒物测量LUC的表达量．结果表明，EPRE调控的LUC报告载体框架正确；LUC的表达量与3种受试物具 有明显的剂量效应关系，细胞对3种受试物敏感性为As 十)Cd2十)Hg2+． 关键词：重金属污染物；虫荧光素酶；基因重组细胞 中图分类号：Xl8 文献标识码：A 文章编号：1000—6923；(2007)06—0835 06 The experimental bioassay for heavy metal pollutants based on the recombinant firefly luciferase gene ceH．LI Xiang-ming ，LUO Fang-ni ，CHEN Chun—bo ，Wu Yu-juarl2，LI Hua-ling (1．Medical College，Yangzhou University, Yangzhou 225001，China；2．College ofAnimal Science and Technology,Yangzhou University,Yangzhou 225001，China： 3．Veterinarian College，Yangzh ， ． ， ． ou University,Yangzhou 225001 China)China Environmental Science 2007(6)：835-840 Abstract：To assess quickly the potential environmental hazard of heavy metal pollutant by the gene recombinant cell，a reporter vector regulated by electrophile response elements(EPRE)Was constructed by fusing EPRE to the TK minimal promoter and the gene encoding firefly luciferase(LUC)with biomolecular techniques．Then this reporter vector Was transiently transformed into HeLa．The recombinant cell Was treated by the different concentration ofNaAsO2，CdC12 and HgC12，the expression of luciferase Was determ ined by luciferase assay kit．The frame of LUC reporter vector regulated by EPRE was correct；the significant dose—dependent relationships were found between the LUC expression and 3 kinds of the test substance concentration．The sensitivity to the cell was as follows：As Cd2 Hg2 ． Key words：heavy metal pollutant；firefly luciferase；