Vazquez-Vilar Marta, Sarrion-Perdigones Alejandro, Ziarsolo Peio, Blanca Jose, Granell Antonio, Orzaez Diego
Instituto de Biología Molecular y Celular de Plantas (IBMCP), Consejo Superior de Investigaciones Científicas, Universidad Politécnica de Valencia, Camino de Vera s/n, 46022, Valencia, Spain.
Methods Mol Biol. 2015;1284:399-420. doi: 10.1007/978-1-4939-2444-8_20.
GoldenBraid (GB) is a modular DNA assembly technology for plant multigene engineering based on type IIS restriction enzymes. GB speeds up the assembly of transcriptional units from standard genetic parts and facilitates the stacking of several genes within the same T-DNA in few days. GBcloning is software-assisted with a set of online tools. The GBDomesticator tool assists in the adaptation of DNA parts to the GBstandard. The combination of GB-adapted parts to build new transcriptional units is assisted by the GB TU Assembler tool. Finally, the assembly of multigene modules is simulated by the GB Binary Assembler. All the software tools are available at www.gbcloning.org . Here, we describe in detail the assembly methodology to create a multigene construct with three transcriptional units for polyphenol metabolic engineering in plants.
GoldenBraid（GB）是一种基于IIS型限制酶的用于植物多基因工程的模块化DNA组装技术。GB加速了从标准遗传元件组装转录单元的过程，并有助于在几天内将多个基因堆叠在同一T-DNA中。GB克隆由一套在线工具提供软件辅助。GBDomesticator工具协助DNA元件适应GB标准。GB TU Assembler工具辅助将适应GB的元件组合以构建新的转录单元。最后，GB Binary Assembler对多基因模块的组装进行模拟。所有软件工具均可在www.gbcloning.org上获取。在此，我们详细描述了用于创建具有三个转录单元的多基因构建体以进行植物多酚代谢工程的组装方法。
