Mori Kotaro, Kashiwagi Akiko, Urabe Itaru, Yomo Tetsuya
Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan.
Biosystems. 2009 Feb;95(2):114-9. doi: 10.1016/j.biosystems.2008.09.001. Epub 2008 Sep 6.
We performed a series of evolution experiments, the results of which illustrated the relationship between mutations and increased carrying capacity (K). Performing an evolution experiment with repeated cycles of mutation by PCR and selection makes it possible to obtain results over shorter culture durations than in methods reported previously relying on spontaneous mutation and selection. We constructed random mutant populations of Escherichia coli in which members differed only in part of the genomic copy of the glutamine synthetase gene and performed daily serial transfer culture where the populations were in K-selected environments. The value of K in this system was increased by 10(5)- to 10(8)-fold relative to the parent clone, which was achieved by four randomly introduced mutations. This method can be applied to any gene and will be useful for analyzing a number of important issues in evolutionary biology.
我们进行了一系列进化实验,实验结果阐明了突变与增加的承载能力(K)之间的关系。通过PCR进行重复的突变循环和选择来开展进化实验,相较于之前报道的依赖自发突变和选择的方法,能够在更短的培养时间内获得结果。我们构建了大肠杆菌的随机突变群体,其中成员仅在谷氨酰胺合成酶基因的部分基因组拷贝上存在差异,并在K选择环境中对群体进行每日连续传代培养。相对于亲本克隆,该系统中的K值增加了10⁵至10⁸倍,这是通过四个随机引入的突变实现的。该方法可应用于任何基因,将有助于分析进化生物学中的许多重要问题。