Mutagenesis is an important technique for microbial mutation breeding.As the source of mutations,DNA damage extent is a key indicator for the effectiveness of mutagenesis.Therefore,a rapid and easy DNA damage quantifi...Mutagenesis is an important technique for microbial mutation breeding.As the source of mutations,DNA damage extent is a key indicator for the effectiveness of mutagenesis.Therefore,a rapid and easy DNA damage quantification method is required for the comparison of mutagenesis effects and development of mutagenesis tools.Here,we used the umu-microplate test system to quantitatively compare the DNA damage strength caused by atmospheric and room-temperature plasma(ARTP)and other traditional mutagenesis methods including:ultraviolet radiation(UV),diethyl sulfate(DES)and 4-nitroquinoline-1-oxide(4-NQO).The test strain of Salmonella typhimurium TA1535/pSK1002 was used to monitor the time-course profile of b-galactosidase activity induced by DNA damage caused by different mutagenesis methods using a microplate reader.The umu-microplate test results showed that ARTP caused higher extent of DNA damage than UV and chemical mutagens,which agrees well with the result obtained by SOS-FACS-based quantification method as reported previously.This umu-microplate test is accessible for broad researchers who are lack of the expensive FACS instruments and allows the quick quantitative evaluation of DNA damage among living cells for different mutagenesis methods in the study of the microbial mutation breeding.展开更多
The genotoxic activities of effluents from drainage water treatment plants were examined by using the novel <em>umu</em> tester strain NM8001, which lacks <em>MutMst</em> genes. To enhance the ...The genotoxic activities of effluents from drainage water treatment plants were examined by using the novel <em>umu</em> tester strain NM8001, which lacks <em>MutMst</em> genes. To enhance the sensitivity of the LacZ assay, a BugBuster mix protein extraction reagent and TokyoGreen-<em>β</em> Gal for a fluorescence-galactosidase substrate were applied. Of the 24 sampling locations present in Kanagawa prefecture, Japan, water extracts from nine sampling points showed apparent genotoxic activities without metabolic activation. In contrast, water extracts from the upper sites of these water treatment plants did not show any significant genotoxic activities. The selected samples with genotoxic activity did not show significant mutagenicity toward Ames strains TA98 and TA100. Genotoxicity was also well correlated with the activity of a classical <em>umu </em>strain of TA1535/pSK1002;these findings indicate that the genotoxicity induced by oxidative damage was not a significant component of the genotoxicity.展开更多
基金National Key Research and Development Project of China(2016YFD0102106)National Key Scientific Instrument and Equipment Project of National Natural Science Foundation of China(21627812).
文摘Mutagenesis is an important technique for microbial mutation breeding.As the source of mutations,DNA damage extent is a key indicator for the effectiveness of mutagenesis.Therefore,a rapid and easy DNA damage quantification method is required for the comparison of mutagenesis effects and development of mutagenesis tools.Here,we used the umu-microplate test system to quantitatively compare the DNA damage strength caused by atmospheric and room-temperature plasma(ARTP)and other traditional mutagenesis methods including:ultraviolet radiation(UV),diethyl sulfate(DES)and 4-nitroquinoline-1-oxide(4-NQO).The test strain of Salmonella typhimurium TA1535/pSK1002 was used to monitor the time-course profile of b-galactosidase activity induced by DNA damage caused by different mutagenesis methods using a microplate reader.The umu-microplate test results showed that ARTP caused higher extent of DNA damage than UV and chemical mutagens,which agrees well with the result obtained by SOS-FACS-based quantification method as reported previously.This umu-microplate test is accessible for broad researchers who are lack of the expensive FACS instruments and allows the quick quantitative evaluation of DNA damage among living cells for different mutagenesis methods in the study of the microbial mutation breeding.
文摘The genotoxic activities of effluents from drainage water treatment plants were examined by using the novel <em>umu</em> tester strain NM8001, which lacks <em>MutMst</em> genes. To enhance the sensitivity of the LacZ assay, a BugBuster mix protein extraction reagent and TokyoGreen-<em>β</em> Gal for a fluorescence-galactosidase substrate were applied. Of the 24 sampling locations present in Kanagawa prefecture, Japan, water extracts from nine sampling points showed apparent genotoxic activities without metabolic activation. In contrast, water extracts from the upper sites of these water treatment plants did not show any significant genotoxic activities. The selected samples with genotoxic activity did not show significant mutagenicity toward Ames strains TA98 and TA100. Genotoxicity was also well correlated with the activity of a classical <em>umu </em>strain of TA1535/pSK1002;these findings indicate that the genotoxicity induced by oxidative damage was not a significant component of the genotoxicity.
