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Development of Combinatorial Technology To Simultaneously Accomplish Detection and Purification of the Heavy Metal Contamination.

开发同时完成重金属污染检测与纯化的组合技术。

基本信息

批准号：
16360267
负责人：
ENDO Ginro
金额：
$ 7.94万
依托单位：
Tohoku Gakuin University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16360267/
关键词：
Low lebel heavy metal contamination Detection and purification Heavy metal biosensor Transgenic yeast ppk gene Transgenic plant vam3gene 水銀汚染浄化技術バイオセンサー複合工学技術重金属

项目摘要

As one of environmental problems in future, low level and wide aerial contamination of heavy metals will be serious for human health. In this study, combinatorial technology for simultaneous detection and purification of the heavy metal contamination was developed. First, sensitive detection system of contaminated heavy metals was developed by introducing heavy metal transportation activity and a reporter gene expression systems into bacterial cells. Second, plants that have heavy metal removal activity were developed using mercury ion transporter gene and polyphosphate synthesis gene, ppk gene, into tobacco cells. Third, specific methods of introduction of a mercury ion transporter gene into yeast cells were investigated to know maldistribution of the gene expression in the yeast cells.The experimental results obtained from the research showed that the developed mercury biosensor has very high sensitivity (with detection limit of 1 nM for inorganic mercury ion and 0.5 nM for organic mercurial compound) and that the gfp fusion biosensor device does not required any special equipment for the mercury detection and it can be applicable for measurement of the mercury contamination in actual environmental sites. From the study using transgenic plant (tobacco), it was understood that ppk gene cloning with a mercury transporter gene, merT, is effective to bread mercury resistant and accumulation plants. Form the study using yeast cell expression system, it was shown that the expressed fusion protein of heavy metal transporter protein, MerC, and a syntaxine family protein, Vam3p is specially located at the vacuole membrane of the yeast cell, and it confers heavy metal resistance and accumulation capability to the yeast cell.

作为未来的环境问题之一，大气中低水平、大范围的重金属污染将严重危害人类健康。本研究开发了同时检测和净化重金属污染的组合技术。首先，通过在细菌细胞中引入重金属转运活性和报告基因表达系统，建立了重金属污染的灵敏检测系统。其次，将汞离子转运蛋白基因和多聚磷酸合成基因PPK基因导入烟草细胞，获得了具有去除重金属活性的植物。第三，研究了将汞离子转运蛋白基因导入酵母细胞的具体方法，以了解该基因在酵母细胞中的表达分布情况。实验结果表明，所研制的汞生物传感器具有很高的灵敏度(对无机汞离子的检测下限为1 nM，对有机汞化合物的检测下限为0.5 nM)，该GFP融合生物传感器装置不需要任何特殊的汞检测设备，可以适用于实际环境场所的汞污染测量。通过利用转基因植物(烟草)进行的研究表明，利用汞转运蛋白基因Mert克隆PPK基因对于培育抗汞和积累汞的植物是有效的。利用酵母细胞表达系统的研究表明，所表达的重金属转运蛋白Merc与合成素家族蛋白Vam3p的融合蛋白定位于酵母细胞的液泡膜上，赋予酵母细胞对重金属的抗性和积累能力。