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

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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.