Streptomyces and other closely-related actinobacteria are important sources of bioactive molecules. Streptomyces synthetic biology and genetics empower therapeutic and agrichemical development through strain improvement and biosynthetic understanding. Such efforts rely on the availability of developed molecular toolsets. Among these tools, vectors that enable combinatorial chromosomal manipulations are particularly desirable. Towards developing tools for facile multiplex engineering, we herein describe the development of new integrating vectors derived from BD1 subgroup actinophage OzzyJ (ϕOZJ). By demonstrating the transformation of several Streptomyces spp. using ϕOZJ-derived vectors, we reveal their potential for strain engineering. We further report the development of new ϕC31 and ϕBT1-based vectors having orthogonal resistance, replication and integration features for concomitant transformation with our ϕOZJ-derived vectors. Importantly, the resulting compatible vector panel enabled us to demonstrate the transfer of up to three plasmids each into Streptomyces venezuelae, Streptomyces roseosporus and Streptomyces pristinaespiralis during a single conjugation experiment. To our knowledge this is the first documentation of conjugation-mediated multiplex plasmid transformation, a useful approach for rapid combinatorial strain development.

中文翻译：

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用act噬菌体ϕOZJ构建新的整合载体，并将其用于多重链霉菌转化。

链霉菌和其他紧密相关的放线菌是生物活性分子的重要来源。链霉菌的合成生物学和遗传学通过菌株改良和生物合成理解来促进治疗和农业化学的发展。这种努力依赖于已开发的分子工具集的可用性。在这些工具中，特别需要能够进行组合染色体操纵的载体。对于用于方便的多重工程的开发工具，我们在这里描述了从BD1子集放线菌OzzyJ（ϕOZJ）派生的新整合载体的开发。通过展示几种链霉菌的变种。使用ϕOZJ衍生的载体，我们揭示了它们在菌株工程中的潜力。我们进一步报告了具有正交抗性的新的基于ϕC31和ϕBT1的载体的开发，复制和整合功能，可与我们的ϕOZJ衍生载体同时转化。重要的是，所得的相容性载体组使我们能够证明在单个缀合实验中将多达三个质粒各自转移至委内瑞斯链霉菌，玫瑰链霉菌和链霉菌链霉菌中。据我们所知，这是结合介导的多重质粒转化的第一个文献，这是快速组合菌株开发的有用方法。