title: Design and construction of multigenic constructs for plant biotechnology using the GoldenBraid cloning strategy.
abstract: 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.
authors.affiliation: Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas (CSIC), Universidad Politécnica de Valencia, Valencia, Spain.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Sarrion-Perdigones
authors.forename: Jorge
authors.initials: J
authors.lastname: Palaci
authors.forename: Antonio
authors.initials: A
authors.lastname: Granell
authors.forename: Diego
authors.initials: D
authors.lastname: Orzaez
mesh_terms: D060054:Agrobacterium / Q000166:cytology / Q000235:genetics; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods*; D018929:Cell Culture Techniques; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics; D005810:Multigene Family / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic