GoldenBraid (GB) is an iterative and standardized DNA assembling system specially designed for Multigene Engineering in Plant Synthetic Biology. GB is based on restriction-ligation reactions using type IIS restriction enzymes. GB comprises a collection of standard DNA pieces named "GB parts" and a set of destination plasmids (pDGBs) that incorporate the multipartite assembly of standardized DNA parts. GB reactions are extremely efficient: two transcriptional units (TUs) can be assembled from several basic GBparts in one T-DNA less than 24 h. Moreover, larger assemblies comprising 4-5 TUs are routinely built in less than 2 working weeks. Here we provide a detailed view of the GB methodology. As a practical example, a Bimolecular Fluorescence Complementation construct comprising four TUs in a 12 kb DNA fragment is presented. © Springer Science+Business Media New York 2014.
CITATION STYLE
Sarrion-Perdigones, A., Palaci, J., Granell, A., & Orzaez, D. (2014). Design and construction of multigenic constructs for Plant biotechnology using the GoldenBraid cloning strategy. Methods in Molecular Biology, 1116, 133–151. https://doi.org/10.1007/978-1-62703-764-8_10
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