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转基因烟草植株中芳烃受体(AhR)介导的报告基因表达系统。

Aryl hydrocarbon receptor (AhR)-mediated reporter gene expression systems in transgenic tobacco plants.

作者信息

Kodama Susumu, Okada Kumiko, Inui Hideyuki, Ohkawa Hideo

机构信息

Graduate School of Science and Technology, Kobe University, Kobe, Hyogo, Japan.

出版信息

Planta. 2007 Dec;227(1):37-45. doi: 10.1007/s00425-007-0592-1. Epub 2007 Sep 19.

Abstract

In mammals, the aryl hydrocarbon receptor (AhR) mediates expression of certain genes, including CYP1A1, in response to exposure to dioxins and related compounds. We have constructed a mouse AhR-mediated gene expression systems for a beta-glucuronidase (GUS) reporter gene consisting of an AhR, an AhR nuclear translocator (Arnt), and a xenobiotic response element (XRE)-driven promoter in transgenic tobacco plants. On treatment with the AhR ligands 3-methylcholanthrene (MC), beta-naphthoflavone (betaNF), and indigo, the transgenic tobacco plants exhibited enhanced GUS activity, presumably by inducible expression of the reporter gene. The recombinant AhR (AhRV), with the activation domain replaced by that of the Herpes simplex virus protein VP16, induced GUS activity much more than the wild-type AhR in the transgenic tobacco plants. Plants carrying AhRV expressed the GUS reporter gene in a dose- and time-dependent manner when treated with MC; GUS activity was detected at 5 nM MC on solid medium and at 12 h after soaking in 25 microM MC. Histochemical GUS staining showed that this system was active mainly in leaf and stem. These results suggest that the AhR-mediated reporter gene expression system has potential for the bioassay of dioxins in the environment and as a novel gene expression system in plants.

摘要

在哺乳动物中,芳烃受体(AhR)介导某些基因(包括CYP1A1)的表达,以响应二噁英及相关化合物的暴露。我们构建了一个用于β-葡萄糖醛酸酶(GUS)报告基因的小鼠AhR介导的基因表达系统,该系统由AhR、AhR核转运蛋白(Arnt)和异生素反应元件(XRE)驱动的启动子组成,并将其转入转基因烟草植株中。在用AhR配体3-甲基胆蒽(MC)、β-萘黄酮(βNF)和靛蓝处理后,转基因烟草植株表现出增强的GUS活性,推测是通过报告基因的诱导表达实现的。重组AhR(AhRV)的激活结构域被单纯疱疹病毒蛋白VP16的激活结构域取代,在转基因烟草植株中诱导GUS活性的能力比野生型AhR强得多。携带AhRV的植株在用MC处理时,GUS报告基因呈剂量和时间依赖性表达;在固体培养基上5 nM的MC处理时以及在25 μM MC浸泡12小时后可检测到GUS活性。组织化学GUS染色表明,该系统主要在叶片和茎中具有活性。这些结果表明,AhR介导的报告基因表达系统在环境中二噁英的生物测定以及作为植物中的一种新型基因表达系统方面具有潜力。

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