The effect of metabolic stress on genome stability of a synthetic biology chassis Escherichia coli K12 strain.
The effect of metabolic stress on genome stability of a synthetic biology chassis Escherichia coli K12 strain.
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代谢应激对合成生物学底盘大肠杆菌K12菌株基因组稳定性的影响。
DOI：
10.1186/s12934-018-0858-2
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发表时间：
2018-01-22
影响因子：
6.4
通讯作者：
Sloan WT
中科院分区：
文献类型：
--
作者：
Couto JM;McGarrity A;Russell J;Sloan WT
越来越多地提出基于合成生物体的生物技术用于环境应用，例如原位传感。通常，这些生物体的新功能是通过编译底盘生物体基因组中的遗传片段来实现的。为了表现出可预测性，这些底盘被设计成具有减少的基因组，从而最小化生物复杂性。然而，在这些提出的应用中，预计即使当包含在装置内时，生物体也将暴露于波动的、通常是应激的条件，并且不清楚它们的基因组是否将保持稳定性。在这里，我们采用恒化器设计，使我们能够保持两个菌株的E。coliK 12在持续饥饿胁迫下的生长：首先是简化基因组合成生物学底盘MDS 42，然后是对照亲本菌株MG 1655。我们估计了突变率，并将其用作基因组不稳定性增加的指标。我们发现，在24小时内，两种菌株的自发突变率增加相似，使基因组不稳定。在实验期间保持高速率。从两个菌株随机取样的突变体的队列的生长速率被用作新兴的表型的代理，并通过关联遗传变异。突变体的生长速率始终低于非突变体的生长速率，这是两种菌株适应性降低和存在轻度有害突变的指标。此外，这些突变对整个种群的影响因菌株而异。总体而言，该研究表明，MDS 42中的基因组减少在代谢应激下不能稳定底盘。随着时间的推移，这可能会损害在环境应用中建立在底盘上的合成生物的有效性。本文的在线版本（10.1186/s12934-018-0858-2）包含补充材料，可供授权用户使用。
Synthetic organism-based biotechnologies are increasingly being proposed for environmental applications, such as in situ sensing. Typically, the novel function of these organisms is delivered by compiling genetic fragments in the genome of a chassis organism. To behave predictably, these chassis are designed with reduced genomes that minimize biological complexity. However, in these proposed applications it is expected that even when contained within a device, organisms will be exposed to fluctuating, often stressful, conditions and it is not clear whether their genomes will retain stability. Here we employed a chemostat design which enabled us to maintained two strains of E. coli K12 under sustained starvation stress: first the reduced genome synthetic biology chassis MDS42 and then, the control parent strain MG1655. We estimated mutation rates and utilised them as indicators of an increase in genome instability. We show that within 24 h the spontaneous mutation rate had increased similarly in both strains, destabilizing the genomes. High rates were maintained for the duration of the experiment. Growth rates of a cohort of randomly sampled mutants from both strains were utilized as a proxy for emerging phenotypic, and by association genetic variation. Mutant growth rates were consistently less than rates in non-mutants, an indicator of reduced fitness and the presence of mildly deleterious mutations in both the strains. In addition, the effect of these mutations on the populations as a whole varied by strain. Overall, this study shows that genome reductions in the MDS42 did not stabilize the chassis under metabolic stress. Over time, this could compromise the effectiveness of synthetic organisms built on chassis in environmental applications. The online version of this article (10.1186/s12934-018-0858-2) contains supplementary material, which is available to authorized users.
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在全球范围内进行生物修复：从试管到地球。
影响因子：
5.7
作者：
de Lorenzo V;Marlière P;Solé R
通讯作者：
Solé R
基因组减少的枯草芽孢杆菌菌株的表征及其在鸟苷和胸苷生产中的应用
影响因子：
6.4
作者：
Li Y;Zhu X;Zhang X;Fu J;Wang Z;Chen T;Zhao X
通讯作者：
Zhao X
影响因子：
10.7
作者：
Karcagi, Ildiko;Draskovits, Gabor;Papp, Balazs
通讯作者：
Papp, Balazs
影响因子：
3.8
作者：
Garibyan, L;Huang, T;Miller, JH
通讯作者：
Miller, JH
DOI：
10.1038/nrg2775
发表时间：
2010-05
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
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