Toward a genetic system in the marine cyanobacterium Prochlorococcus.
Toward a genetic system in the marine cyanobacterium Prochlorococcus.
复制标题
迈向海洋蓝细菌核酸菌群中的遗传系统。
DOI：
10.1099/acmi.0.000107
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发表时间：
2020
影响因子：
--
通讯作者：
Chisholm SW
中科院分区：
文献类型：
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作者：
Laurenceau R;Bliem C;Osburne MS;Becker JW;Biller SJ;Cubillos-Ruiz A;Chisholm SW
作为海洋中最小和最丰富的初级生产者，蓝细菌原绿球藻引起了不同科学分支的兴趣。在过去的30年里，对这种最小的光养生物的研究使人们对从基因组到全球海洋生态系统的多个尺度的生物组织有了越来越多的了解。在理解其多样性和生态学的驱动因素以及支持其简化简单性的分子机制方面的进展，由于无法从遗传上操纵这些细胞而受到阻碍。多年来，已经进行了多次尝试以开发用于原绿球藻的有效遗传转化方法;迄今为止，所有这些尝试都没有成功，尽管它们的近亲聚球藻取得了一些成功。为了避免追求非生产性的路径，我们在这里报告了我们手中没有工作的方法，以及我们开发一种方法来筛选最有效的电穿孔参数以将最佳DNA递送到原绿球藻细胞中的进展。我们还报告了一种新的协议，获得无菌菌落和区分活细胞和死细胞的新方法。电穿孔方法可用于优化DNA递送到任何细菌中，使其成为用于推进其他遗传转化微生物中的转化系统的有用工具。
As the smallest and most abundant primary producer in the oceans, the cyanobacterium Prochlorococcus is of interest to diverse branches of science. For the past 30 years, research on this minimal phototroph has led to a growing understanding of biological organization across multiple scales, from the genome to the global ocean ecosystem. Progress in understanding drivers of its diversity and ecology, as well as molecular mechanisms underpinning its streamlined simplicity, has been hampered by the inability to manipulate these cells genetically. Multiple attempts have been made to develop an efficient genetic transformation method for Prochlorococcus over the years; all have been unsuccessful to date, despite some success with their close relative, Synechococcus . To avoid the pursuit of unproductive paths, we report here what has not worked in our hands, as well as our progress developing a method to screen the most efficient electroporation parameters for optimal DNA delivery into Prochlorococcus cells. We also report a novel protocol for obtaining axenic colonies and a new method for differentiating live and dead cells. The electroporation method can be used to optimize DNA delivery into any bacterium, making it a useful tool for advancing transformation systems in other genetically recalcitrant microorganisms.