title: Software-assisted stacking of gene modules using GoldenBraid 2.0 DNA-assembly framework.
abstract: 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.
authors.affiliation: 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.
authors.forename: Marta
authors.initials: M
authors.lastname: Vazquez-Vilar
authors.forename: Alejandro
authors.initials: A
authors.lastname: Sarrion-Perdigones
authors.forename: Peio
authors.initials: P
authors.lastname: Ziarsolo
authors.forename: Jose
authors.initials: J
authors.lastname: Blanca
authors.forename: Antonio
authors.initials: A
authors.lastname: Granell
authors.forename: Diego
authors.initials: D
authors.lastname: Orzaez
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004269:DNA, Bacterial; D005818:Genetic Engineering / Q000379:methods*; D023281:Genomics / Q000379:methods*; D020407:Internet; D012984:Software
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA