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Adaptive Evolution of Escherichia coli K-12 MG1655 during Growth on a Nonnative Carbon Source, l-1,2-Propanediol▿ †

机译:大肠杆菌K-12 MG1655在非天然碳源l-1,2-丙二醇上的适应性进化†

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摘要

Laboratory adaptive evolution studies can provide key information to address a wide range of issues in evolutionary biology. Such studies have been limited thus far by the inability of workers to readily detect mutations in evolved microbial strains on a genome scale. This limitation has now been overcome by recently developed genome sequencing technology that allows workers to identify all accumulated mutations that appear during laboratory adaptive evolution. In this study, we evolved Escherichia coli K-12 MG1655 with a nonnative carbon source, l-1,2-propanediol (l-1,2-PDO), for ∼700 generations. We found that (i) experimental evolution of E. coli for ∼700 generations in 1,2-PDO-supplemented minimal medium resulted in acquisition of the ability to use l-1,2-PDO as a sole carbon and energy source so that the organism changed from an organism that did not grow at all initially to an organism that had a growth rate of 0.35 h−1; (ii) six mutations detected by whole-genome resequencing accumulated in the evolved E. coli mutant over the course of adaptive evolution on l-1,2-PDO; (iii) five of the six mutations were within coding regions, and IS5 was inserted between two fuc regulons; (iv) two major mutations (mutations in fucO and its promoter) involved in l-1,2-PDO catabolism appeared early during adaptive evolution; and (v) multiple defined knock-in mutant strains with all of the mutations had growth rates essentially matching that of the evolved strain. These results provide insight into the genetic basis underlying microbial evolution for growth on a nonnative substrate.
机译:实验室适应性进化研究可以提供关键信息,以解决进化生物学中的各种问题。迄今为止,由于工人无法在基因组规模上容易地检测到进化出的微生物菌株中的突变,这种研究受到了限制。现在,通过最近开发的基因组测序技术已经克服了这一局限,该技术使工作人员能够识别在实验室适应性进化过程中出现的所有累积突变。在这项研究中,我们进化了具有非天然碳源l-1,1,2-丙二醇(l,1,2-PDO)的大肠杆菌K-12 MG1655约700代。我们发现(i)在添加1,2-PDO的基本培养基中进行约700代大肠杆菌的实验进化导致获得了使用I-1,2-PDO作为唯一碳和能源的能力,因此该生物从最初完全不生长的生物变为生长速率为0.35 h-1的生物; (ii)在1-1,2-PDO的适应性进化过程中,通过全基因组重测序检测到的六种突变积累在进化的大肠杆菌突变体中; (iii)六个突变中的五个在编码区内,且IS5插入两个fuc调控子之间; (iv)参与l-1,2-PDO分解代谢的两个主要突变(fucO及其启动子突变)出现在适应性进化的早期; (v)具有所有突变的多个确定的敲入突变体菌株的生长速率基本上与进化菌株的生长速率匹配。这些结果为深入了解微生物在非天然底物上生长的遗传基础提供了见识。

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