Cooke Matthew B, Welch Kobie T, Ramirez Laura D, Wen Alice X, Marciano David C, Herman Christophe
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, United States.
Synth Biol (Oxf). 2025 Aug 14;10(1):ysaf015. doi: 10.1093/synbio/ysaf015. eCollection 2025.
Modular cloning systems streamline laboratory workflows by consolidating genetic 'parts' into reusable and modular collections, enabling researchers to fast-track strain construction. The GoldenBraid 2.0 modular cloning system utilizes the cutting property of type IIS restriction enzymes to create defined genetic 'grammars', which facilitate the reuse of standardized genetic parts and assembly of genetic parts in the right order. Here, we present a GoldenBraid 2.0 toolkit of genetic parts designed to accelerate cloning in the model bacterium . This toolkit features 478 pre-made parts for gene expression and protein tagging as well as strains to expedite cloning and strain construction, enabling researchers to quickly generate functional plasmid-borne or chromosome-integrated expression constructs. In addition, we provide a complete laboratory manual with overviews of common reagent recipes, protocols, and community resources to promote toolkit utilization. By streamlining the assembly process, this resource will reduce the financial and temporal burdens of cloning and strain building in many laboratory settings.
模块化克隆系统通过将基因“元件”整合到可重复使用的模块化集合中，简化了实验室工作流程，使研究人员能够快速构建菌株。GoldenBraid 2.0模块化克隆系统利用IIS型限制酶的切割特性来创建特定的基因“语法”，这有助于标准化基因元件的重复使用以及基因元件按正确顺序进行组装。在此，我们展示了一个旨在加速在模式细菌中进行克隆的GoldenBraid 2.0基因元件工具包。该工具包具有478个用于基因表达和蛋白质标签的预制元件以及用于加快克隆和菌株构建的菌株，使研究人员能够快速生成功能性的质粒携带或染色体整合表达构建体。此外，我们提供了一本完整的实验室手册，其中概述了常见试剂配方、方案和社区资源，以促进工具包的使用。通过简化组装过程，该资源将减轻许多实验室环境中克隆和菌株构建的财务和时间负担。
