Kodama Susumu, Okada Kumiko, Akimoto Keiko, Inui Hideyuki, Ohkawa Hideo
Graduate School of Science and Technology, Kobe University, Rokkodai-cho 1-1, Nada-ku, Kobe 657-8501, Japan.
Plant Biotechnol J. 2009 Feb;7(2):119-28. doi: 10.1111/j.1467-7652.2008.00378.x. Epub 2008 Nov 26.
Dioxin residues widely contaminate soil and agricultural products at low concentrations and may accumulate in organisms at the top of food chains owing to their physicochemical properties. In this study, we have developed novel, dioxin-inducible, reporter gene expression systems regulated by recombinant aryl hydrocarbon receptors (AhRs). The recombinant AhRs, referred to as XDVs, consist of the DNA-binding domain of the bacterial repressor protein LexA, a 90-kDa heat shock protein- and ligand-binding regulatory domain from mouse AhR, and the transactivation domain of herpes simplex virus regulatory protein VP16. Transgenic tobacco plants carrying XDVs absorb various AhR ligands, including 3-methylcholanthrene, beta-naphthoflavone and indigo from solid medium and vermiculite, and show dose- and time-dependent expression of the beta-glucuronidase reporter gene. The results clearly suggest that XDVs are functional transcription factors that respond to AhR ligands, and that the XDV-mediated reporter gene expression system is applicable to bioassays for dioxin residues in the environment.
二噁英残留以低浓度广泛污染土壤和农产品,由于其物理化学性质,可能在食物链顶端的生物体中积累。在本研究中,我们开发了由重组芳烃受体(AhR)调控的新型二噁英诱导型报告基因表达系统。重组AhR称为XDVs,由细菌阻遏蛋白LexA的DNA结合结构域、来自小鼠AhR的90 kDa热休克蛋白和配体结合调节结构域以及单纯疱疹病毒调节蛋白VP16的反式激活结构域组成。携带XDVs的转基因烟草植物从固体培养基和蛭石中吸收各种AhR配体,包括3-甲基胆蒽、β-萘黄酮和靛蓝,并显示β-葡萄糖醛酸酶报告基因的剂量和时间依赖性表达。结果清楚地表明,XDVs是对AhR配体有反应的功能性转录因子,并且XDV介导的报告基因表达系统适用于环境中二噁英残留的生物测定。