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Applied Microbiology and Biotechnology

, Volume 99, Issue 13, pp 5639–5646 | Cite as

Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and conventional mutagenesis

  • Xue Zhang
  • Chong ZhangEmail author
  • Qian-Qian Zhou
  • Xiao-Fei Zhang
  • Li-Yan Wang
  • Hai-Bo Chang
  • He-Ping Li
  • Yoshimitsu Oda
  • Xin-Hui XingEmail author
Methods and protocols

Abstract

DNA damage is the dominant source of mutation, which is the driving force of evolution. Therefore, it is important to quantitatively analyze the DNA damage caused by different mutagenesis methods, the subsequent mutation rates, and their relationship. Atmospheric and room temperature plasma (ARTP) mutagenesis has been used for the mutation breeding of more than 40 microorganisms. However, ARTP mutagenesis has not been quantitatively compared with conventional mutation methods. In this study, the umu test using a flow-cytometric analysis was developed to quantify the DNA damage in individual viable cells using Salmonella typhimurium NM2009 as the model strain and to determine the mutation rate. The newly developed method was used to evaluate four different mutagenesis systems: a new ARTP tool, ultraviolet radiation, 4-nitroquinoline-1-oxide (4-NQO), and N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) mutagenesis. The mutation rate was proportional to the corresponding SOS response induced by DNA damage. ARTP caused greater DNA damage to individual living cells than the other conventional mutagenesis methods, and the mutation rate was also higher. By quantitatively comparing the DNA damage and consequent mutation rate after different types of mutagenesis, we have shown that ARTP is a potentially powerful mutagenesis tool with which to improve the characteristics of microbial cell factories.

Keywords

ARTP mutagenesis Flow cytometry Fluctuation test umu test SOS response 

Notes

Acknowledgments

This work was supported by the Tsinghua University Initiative Scientific Research Program (2011THZ 01019) and the JST CREST project of Japan.

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Xue Zhang
    • 1
  • Chong Zhang
    • 1
    • 2
    Email author
  • Qian-Qian Zhou
    • 2
  • Xiao-Fei Zhang
    • 3
  • Li-Yan Wang
    • 1
    • 2
  • Hai-Bo Chang
    • 1
  • He-Ping Li
    • 2
    • 3
  • Yoshimitsu Oda
    • 4
  • Xin-Hui Xing
    • 1
    • 2
    Email author
  1. 1.Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Department of Chemical EngineeringTsinghua UniversityBeijingChina
  2. 2.Biobreeding Research Center, Wuxi Research Institute of Applied TechnologiesTsinghua UniversityWuxiChina
  3. 3.Department of Engineering PhysicsTsinghua UniversityBeijingChina
  4. 4.Institute of Life and Environmental SciencesOsaka Shin-Ai CollegeOsakaJapan

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