Non-model microorganisms often possess complex phenotypes that could be important for the future of biofuel and chemical production. They have received significant interest the last several years, but advancement is still slow due to the lack of a robust genetic toolbox in most organisms. Typically, “domestication” of a new non-model microorganism has been done on an ad hoc basis, and historically, it can take years to develop transformation and basic genetic tools. Here, we review the barriers and solutions to rapid development of genetic transformation tools in new hosts, with a major focus on Restriction-Modification systems, which are a well-known and significant barrier to efficient transformation. We further explore the tools and approaches used for efficient gene deletion, DNA insertion, and heterologous gene expression. Finally, more advanced and high-throughput tools are now being developed in diverse non-model microbes, paving the way for rapid and multiplexed genome engineering for biotechnology.

中文翻译：

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快速驯化非模式微生物的遗传工具开发方法

非模型微生物通常具有复杂的表型，这可能对未来生物燃料和化学生产很重要。在过去的几年中，它们引起了人们的极大兴趣，但是由于大多数生物中缺乏强大的遗传工具箱，因此进展仍然缓慢。通常，新的非模型微生物的“驯化”是临时完成的，从历史上看，开发转化和基本的遗传工具可能要花费数年的时间。在这里，我们回顾了在新宿主中快速发展基因转化工具的障碍和解决方案，主要关注限制-修饰系统，这是有效转化的众所周知且重要的障碍。我们进一步探索了用于有效基因删除，DNA插入和异源基因表达的工具和方法。最后，