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SOS/ umu -Test as A Tool for Genotoxicity Assessment

SOS/ umu -Test as A Tool for Genotoxicity Assessment. By 劉文堅. Advisor: 袁如馨. 防災與水環境研究中心. Why umu -test ?. A low cost experiment assay : no high cost chemicals and complicated apparatus is needed in conducting the experiment.

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SOS/ umu -Test as A Tool for Genotoxicity Assessment

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  1. SOS/umu-Test as A Tool for Genotoxicity Assessment By 劉文堅 Advisor: 袁如馨 防災與水環境研究中心

  2. Why umu-test ? • A low cost experiment assay: no high cost chemicals and complicated apparatus is needed in conducting the experiment. • High-throughput: umu-test enable high amount of sample to be tested at a time. • Short time requirement: umu-test utilizes bacteria which has short life cycle. • High sensitivity: accuracy of umu-test has been proven

  3. Theory of umu-test Mutagen exposure • Firstly, Salmonella typhimuium TA1535/pKK1002 will be exposed to genotoxicants • As soon as the bacteria senses genotoxicity existance, SOS response is transmitted to produce umuC-LacZ fused protein • The protein will produce β-galactosidase • ONPG is used as color indicator • ONPG (colorless) will be converted into o-nitrophenyl (yellow color) by the existance of β-galactosidase • Absorbance measurement is done under spectrophotometer • The amount of β-galactosidasecorrelates to the level of toxicity • Induction ratio obtained by calculation Sos response translation umuC-LacZ fused protein Enzyme production β-galactosidase o-nitrophenyl ONPG (colourless) (Yellow-colored) Absorbance measurement IR calculation

  4. Genotoxicity test of 4-NQO The recent conducted toxicity test result on 4-NQO (well known genotoxicants) indicates the ability of umu test in toxic assessment. The Dashed line represent concentration of 4-NQO at 2-fold greater than the control response, which tells us every black point (IR point) that is higher than the dashed line are considered to be genotoxic.

  5. Future Study Application of umu-test on various toxic samples in the environment. Continuous cooperation with the inventor of the umu-test to increase the efficiency of the test by experimenting on different strains of Salmonella. Comparison of the genotoxicity result by umu-test with those by Comet assay, to establish the database of potential genotoxicants from industrial wastewater.

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