二恶英反应增强子调控的虫荧光素酶报告基因质粒的构建

Construction of Luciferase Reporter Plasmid Which is Under the Control of Dioxin-responsive Enhancers

为加强二恶英类化学物质的快速筛选和半定量检测 ,我们构建了一在二恶英反应增强子调控下的虫荧光素酶报告基因质粒。二恶英反应增强子来源于pHAV质粒 ,MMTV启动子来源于 pCatM质粒 ,上述两者连接后与虫荧光素酶载体连接 ,转染人HepG2肝癌细胞 ,以 2 ,3,7,8 四氯代二苯并二恶英 (TCDD)诱导报告基因表达后检测虫荧光素酶活性。结果表明该质粒中虫荧光素酶的表达受二恶英反应增强子的调控 ,且在一定浓度范围内虫荧光素酶的活性与TCDD的量呈线性关系。研究显示该质粒转染的细胞株有望用于快速筛选及半定量检测二恶英 ,可进一步加强研究作为二恶英类化学物质监测的常规方法。

To improve the bio\|detection of dioxin\|type chemicals,we constructed a plasmid containing reporter under the control of dioxin\|responsive enhancer.After connected the enhancer from the plasmid pHAV with the MMTV promoter of the plasmid pCatM,the connected segment was linked with the luciferase vector.Then the recombinant plasmid was transfected to the HepG2 human hepatoma cell line,and the luciferase activity was induced by 2,3,7,8\|TCDD.The result suggested that expression of the recombinant luciferase was controlled by the dioxin\|responsive enhancer,and there were a linear relationship between the activity of luciferase and certain range of 2,3,7,8\|TCDD concentration.It is promising that the transfected cell line could be used for rapid screen and semi quantitation of dioxin\|type chemicals,and a standard and routine method for the detection of dioxin\|type chemicals by this recombinant plasmid is being under way.