| Budget Amount *help |
¥15,990,000 (Direct Cost: ¥12,300,000、Indirect Cost: ¥3,690,000)
Fiscal Year 2020: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2019: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2018: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
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| Outline of Final Research Achievements |
We have developed a wide variety of bioassay yeasts to detect trace contaminants. In this project, we digested/disrupted the yeast cell wall to increase contaminants influx, and disrupted the drug efflux pump gene to keep the contaminants in the cell. Cell wall mannan protein genes Cwp1, Cwp2, drug efflux pumps Pdr5, Pdr10, Pdr11, Pdr12, Snq2, Yor1, Ste1, and Aus1 were disrupted with the Cre-loxP method for 12 yeast strains. As a result, sensitivities of 8 receptor ligand activity detection strains (dioxin receptor, estrogen receptor alpha, estrogen receptor beta, androgen receptor, progesterone receptor, glucocorticoid receptor, mineralocorticoid receptor, and vitamin D receptor) were improved with mainly combined disruption of Cwp1, Cwp2, Pdr5, and Pdr10 genes. In some case, yeast protoplast after enzymatic cell wall digestion showed higher performance. Unexpectedly, there are strains of which sensitivity were not improved the at all, so further sturdy should be required.
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