1 Center for Biosystems Dynamics Research (BDR), RIKEN, Osaka, Japan.
2 Universal Biology Institute, School of Science, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
SLAS Technol. 2019 Jun;24(3):342-348. doi: 10.1177/2472630318800283. Epub 2018 Sep 10.
Ultraviolet (UV) mutagenesis is a widely used technique to increase bacterial mutation rates in laboratory experiments. UV mutagenesis requires fine regulation of UV dose, because the number of dead cells increases exponentially as the dose increases. Ignoring this hazard can cause extinction of UV-exposed populations. Therefore, an automated system that cooperatively conducts both growth measurement and UV irradiation is needed for efficient UV mutagenesis experiments. To address this task, we constructed an automated UV irradiation device for microbial cell culture. This device can measure cell density and irradiate the bacterial cells with UV light automatically according to the state of cell growth. We demonstrated that this growth feedback control avoided extinction and enabled accumulation of mutations in bacterial genomes at a rapid rate for a long period. Whole-genome sequencing revealed the high accumulation rate, neutrality, and spectrum of UV-induced mutations. These characteristics were all consistent with those obtained by manual UV irradiation. These results indicate that our automated device is useful in accelerating mutation accumulation over a long duration.
紫外线(UV)诱变是一种广泛应用于实验室实验的增加细菌突变率的技术。UV 诱变需要对 UV 剂量进行精细调节,因为随着剂量的增加,死亡细胞的数量呈指数级增加。忽略这一危险可能导致 UV 暴露的种群灭绝。因此,需要一种能够协同进行生长测量和 UV 照射的自动化系统,以进行高效的 UV 诱变实验。为了解决这个问题,我们构建了一种用于微生物细胞培养的自动化 UV 照射装置。该装置可以根据细胞生长状态自动测量细胞密度并对细菌细胞进行 UV 照射。我们证明,这种生长反馈控制避免了灭绝,并使细菌基因组中的突变以较快的速度迅速积累。全基因组测序揭示了 UV 诱导突变的高积累率、中性和频谱。这些特征与手动 UV 照射获得的结果一致。这些结果表明,我们的自动化装置可用于在较长时间内加速突变积累。