Institute of Parasitology, Biology Centre, Czech Academy of Sciences and Faculty of Sciences, University of South Bohemia, České Budějovice, Czech Republic.
Department of Biochemistry, University of Cambridge, Cambridge, UK.
Nat Methods. 2020 May;17(5):481-494. doi: 10.1038/s41592-020-0796-x. Epub 2020 Apr 6.
Diverse microbial ecosystems underpin life in the sea. Among these microbes are many unicellular eukaryotes that span the diversity of the eukaryotic tree of life. However, genetic tractability has been limited to a few species, which do not represent eukaryotic diversity or environmentally relevant taxa. Here, we report on the development of genetic tools in a range of protists primarily from marine environments. We present evidence for foreign DNA delivery and expression in 13 species never before transformed and for advancement of tools for eight other species, as well as potential reasons for why transformation of yet another 17 species tested was not achieved. Our resource in genetic manipulation will provide insights into the ancestral eukaryotic lifeforms, general eukaryote cell biology, protein diversification and the evolution of cellular pathways.
多样的微生物生态系统是海洋生命的基础。这些微生物中有许多单细胞真核生物,它们跨越了真核生物生命树的多样性。然而,遗传的可操作性一直局限于少数几个物种,这些物种不能代表真核生物的多样性或具有环境相关性的分类群。在这里,我们报告了一系列主要来自海洋环境的原生生物的遗传工具的开发。我们提供了证据证明 13 个以前从未转化过的物种可以进行外源 DNA 的传递和表达,以及另外 8 个物种的工具的改进,以及为什么对另外 17 个测试的物种没有实现转化的潜在原因。我们在遗传操作方面的资源将为研究原始真核生物形态、一般真核生物细胞生物学、蛋白质多样化以及细胞途径的进化提供新的见解。