Quantitative evaluation of DNA damage caused by atmospheric and room-temperature plasma (ARTP) and other mutagenesis methods using a rapid umu-microplate test protocol for microbial mutation breeding

doi:10.1016/j.cjche.2021.01.009

中国化学工程学报 ›› 2021, Vol. 39 ›› Issue (11): 205-210.DOI: 10.1016/j.cjche.2021.01.009

• Biotechnology and Bioengineering • 上一篇下一篇

Quantitative evaluation of DNA damage caused by atmospheric and room-temperature plasma (ARTP) and other mutagenesis methods using a rapid umu-microplate test protocol for microbial mutation breeding

Yuting Huang¹, Liyang Wang², Xue Zhang³, Nan Su¹, Heping Li⁴, Yoshimitsu Oda⁵, Xinhui Xing^1,6,7,8

1 Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China;
2 Biobreeding Research Center, Wuxi Research Institute of Applied Technologies, Tsinghua University, Wuxi 214072, China;
3 Department of Entomology, College of Plant Protection, China Agricultural University, Beijing 100193, China;
4 Department of Engineering Physics, Tsinghua University, Beijing 100084, China;
5 Institute of Life and Environmental Sciences, Osaka Shin-Ai College, Tsurumi, Tsurumi-ku, Osaka 5380052, Japan;
6 Centre for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China;
7 Institute of Biopharmaceutical and Health Engineering, Tsinghua Shenzhen International Graduate School, Shenzhen 518055, China;
8 Institute of Biomedical Health Technology and Engineering, Shenzhen Bay Laboratory, Shenzhen 518055, China

收稿日期:2020-09-28 修回日期:2021-01-22 出版日期:2021-11-28 发布日期:2021-12-27
通讯作者: Xinhui Xing
基金资助:
This work was supported by National Key Research and Development Project of China (2016YFD0102106) and National Key Scientific Instrument and Equipment Project of National Natural Science Foundation of China (21627812).

Quantitative evaluation of DNA damage caused by atmospheric and room-temperature plasma (ARTP) and other mutagenesis methods using a rapid umu-microplate test protocol for microbial mutation breeding

Yuting Huang¹, Liyang Wang², Xue Zhang³, Nan Su¹, Heping Li⁴, Yoshimitsu Oda⁵, Xinhui Xing^1,6,7,8

1 Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China;
2 Biobreeding Research Center, Wuxi Research Institute of Applied Technologies, Tsinghua University, Wuxi 214072, China;
3 Department of Entomology, College of Plant Protection, China Agricultural University, Beijing 100193, China;
4 Department of Engineering Physics, Tsinghua University, Beijing 100084, China;
5 Institute of Life and Environmental Sciences, Osaka Shin-Ai College, Tsurumi, Tsurumi-ku, Osaka 5380052, Japan;
6 Centre for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China;
7 Institute of Biopharmaceutical and Health Engineering, Tsinghua Shenzhen International Graduate School, Shenzhen 518055, China;
8 Institute of Biomedical Health Technology and Engineering, Shenzhen Bay Laboratory, Shenzhen 518055, China

Received:2020-09-28 Revised:2021-01-22 Online:2021-11-28 Published:2021-12-27
Contact: Xinhui Xing
Supported by:
This work was supported by National Key Research and Development Project of China (2016YFD0102106) and National Key Scientific Instrument and Equipment Project of National Natural Science Foundation of China (21627812).

摘要/Abstract

摘要： 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 β-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.

关键词: ARTP mutagenesis, umu-Microplate test, Biological engineering, Cell engineering, Biotechnology

Abstract: 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 β-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.

Key words: ARTP mutagenesis, umu-Microplate test, Biological engineering, Cell engineering, Biotechnology

Yuting Huang, Liyang Wang, Xue Zhang, Nan Su, Heping Li, Yoshimitsu Oda, Xinhui Xing. Quantitative evaluation of DNA damage caused by atmospheric and room-temperature plasma (ARTP) and other mutagenesis methods using a rapid umu-microplate test protocol for microbial mutation breeding[J]. 中国化学工程学报, 2021, 39(11): 205-210.

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Quantitative evaluation of DNA damage caused by atmospheric and room-temperature plasma (ARTP) and other mutagenesis methods using a rapid umu-microplate test protocol for microbial mutation breeding

Quantitative evaluation of DNA damage caused by atmospheric and room-temperature plasma (ARTP) and other mutagenesis methods using a rapid umu-microplate test protocol for microbial mutation breeding

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