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线粒体基因组左臂的化学合成及功能分析

Chemical synthesis of left arm of mitochondrial genome and functional analysis.

作者信息

Wang Quan, Luo Haolin, Zhuang Jieyi, Li Xinyi, Huang Danqiong, Hu Zhangli, Zhang Guiying

机构信息

Guangdong Technology Research Center for Marine Algal Bioengineering, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, China.

Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic Engineering, Shenzhen University, Shenzhen, China.

出版信息

Front Microbiol. 2022 Dec 22;13:1064497. doi: 10.3389/fmicb.2022.1064497. eCollection 2022.

Abstract

is a photosynthetic eukaryote showing great industrial potential. The synthesis and function of the artificial genome not only promotes the development of synthetic biology technology but also supports industries that utilize this algae. Mitochondrial genome (MtG) is the smallest and simplest genome of that suits synthetic exploration. In this article, we designed and assembled a synthetic mitochondria left arm (syn-LA) genome sharing >92% similarity to the original mitochondria genome (OMtG) left arm, transferred it into the respiratory defect strain , screened syn-LA containing transformants from recovered dark-growth defects using PCR amplification, verified internal function of syn-LA western blot, detected heteroplasmic ratio of syn-LA, tried promoting syn-LA into homoplasmic status with paromomycin stress, and discussed the main limitations and potential solutions for this area of research. This research supports the functionalization of a synthetic mitochondrial genome in living cells. Although further research is needed, this article nevertheless provides valuable guidance for the synthesis of eukaryotic organelle genomes and opens possible directions for future research.

摘要

是一种具有巨大工业潜力的光合真核生物。人工基因组的合成与功能不仅推动了合成生物学技术的发展,也为利用这种藻类的产业提供了支持。线粒体基因组(MtG)是最适合进行合成探索的最小且最简单的基因组。在本文中,我们设计并组装了一个与原始线粒体基因组(OMtG)左臂相似度大于92%的合成线粒体左臂(syn-LA)基因组,将其转入呼吸缺陷菌株,通过PCR扩增从恢复的暗生长缺陷中筛选出含有syn-LA的转化体,通过蛋白质免疫印迹验证syn-LA的内部功能,检测syn-LA的异质性比率,尝试用巴龙霉素胁迫促使syn-LA达到同质性状态,并讨论了该研究领域的主要局限性和潜在解决方案。本研究支持合成线粒体基因组在活细胞中的功能化。尽管还需要进一步研究,但本文仍然为真核细胞器基因组的合成提供了有价值的指导,并为未来的研究开辟了可能的方向。

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