title: Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and conventional mutagenesis.
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.
authors.affiliation: Key Laboratory for Industrial Biocatalysis, Ministry of Education of China, Department of Chemical Engineering, Tsinghua University, Beijing, China.
authors.forename: Xue
authors.initials: X
authors.lastname: Zhang
authors.forename: Chong
authors.initials: C
authors.lastname: Zhang
authors.forename: Qian-Qian
authors.initials: QQ
authors.lastname: Zhou
authors.forename: Xiao-Fei
authors.initials: XF
authors.lastname: Zhang
authors.forename: Li-Yan
authors.initials: LY
authors.lastname: Wang
authors.forename: Hai-Bo
authors.initials: HB
authors.lastname: Chang
authors.forename: He-Ping
authors.initials: HP
authors.lastname: Li
authors.forename: Yoshimitsu
authors.initials: Y
authors.lastname: Oda
authors.forename: Xin-Hui
authors.initials: XH
authors.lastname: Xing
mesh_terms: D004249:DNA Damage / Q000187:drug effects* / Q000528:radiation effects*; D005434:Flow Cytometry; D016296:Mutagenesis; D059645:Mutation Rate; D058626:Plasma Gases; D012486:Salmonella typhimurium / Q000187:drug effects* / Q000528:radiation effects*; D014466:Ultraviolet Rays
chemical_list: D058626:Plasma Gases