Cochrane Ryan R, Brumwell Stephanie L, Shrestha Arina, Giguere Daniel J, Hamadache Samir, Gloor Gregory B, Edgell David R, Karas Bogumil J
Department of Biochemistry, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON N6A 5C1, Canada.
Biology (Basel). 2020 Oct 26;9(11):358. doi: 10.3390/biology9110358.
Algae are attractive organisms for biotechnology applications such as the production of biofuels, medicines, and other high-value compounds due to their genetic diversity, varied physical characteristics, and metabolic processes. As new species are being domesticated, rapid nuclear and organelle genome engineering methods need to be developed or optimized. To that end, we have previously demonstrated that the mitochondrial genome of microalgae can be cloned and engineered in and . Here, we show that the same approach can be used to clone mitochondrial genomes of another microalga, We have demonstrated that these genomes can be cloned in as easily as those of , but they are less stable when propagated in . Specifically, after approximately 60 generations of propagation in , 17% of cloned mitochondrial genomes contained deletions compared to 0% of previously cloned mitochondrial genomes. This genome instability is potentially due to the lower G+C DNA content of (30%) compared to (35%). Consequently, the previously established method can be applied to clone 's mitochondrial genome, however, more frequent analyses of genome integrity will be required following propagation in prior to use in downstream applications.
由于藻类具有遗传多样性、多样的物理特性和代谢过程,它们是生物技术应用(如生物燃料、药物和其他高价值化合物的生产)中具有吸引力的生物体。随着新物种被驯化,需要开发或优化快速的核基因组和细胞器基因组工程方法。为此,我们之前已经证明微藻的线粒体基因组可以在[具体条件1]和[具体条件2]中进行克隆和工程改造。在这里,我们表明相同的方法可用于克隆另一种微藻的线粒体基因组,我们已经证明这些基因组可以像[第一种微藻]的基因组一样容易地在[具体条件3]中克隆,但在[具体条件4]中繁殖时它们的稳定性较差。具体而言,在[具体条件4]中繁殖约60代后,17%的克隆[第二种微藻]线粒体基因组包含缺失,而之前克隆的[第一种微藻]线粒体基因组的缺失率为0%。这种基因组不稳定性可能是由于[第二种微藻]的G+C DNA含量(30%)低于[第一种微藻](35%)。因此,先前建立的方法可用于克隆[第二种微藻]的线粒体基因组,然而,在用于下游应用之前,在[具体条件4]中繁殖后需要更频繁地分析基因组完整性。