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Title

GoldenBraid: An iterative cloning system for standardized assembly of reusable genetic modules

AuthorsSarrion-Perdigones, Alejandro ; Elvira Falconi, Erica; Zandalinas, Sara I.; Juárez, Paloma ; Fernández-del-Carmen, Asun ; Granell, Antonio ; Orzáez, Diego
KeywordsMediated plant transformation
One-pot
Expression
Transcription
Issue Date7-Jul-2011
PublisherPublic Library of Science
CitationPlos One 6(7): e21622 (2011)
AbstractSynthetic Biology requires efficient and versatile DNA assembly systems to facilitate the building of new genetic modules/ pathways from basic DNA parts in a standardized way. Here we present GoldenBraid (GB), a standardized assembly system based on type IIS restriction enzymes that allows the indefinite growth of reusable gene modules made of standardized DNA pieces. The GB system consists of a set of four destination plasmids (pDGBs) designed to incorporate multipartite assemblies made of standard DNA parts and to combine them binarily to build increasingly complex multigene constructs. The relative position of type IIS restriction sites inside pDGB vectors introduces a double loop (‘‘braid’’) topology in the cloning strategy that allows the indefinite growth of composite parts through the succession of iterative assembling steps, while the overall simplicity of the system is maintained. We propose the use of GoldenBraid as an assembly standard for Plant Synthetic Biology. For this purpose we have GB-adapted a set of binary plasmids for A. tumefaciens-mediated plant transformation. Fast GB-engineering of several multigene T-DNAs, including two alternative modules made of five reusable devices each, and comprising a total of 19 basic parts are also described.
DescriptionThis is an open-access article distributed under the terms of the Creative Commons Attribution License.
Publisher version (URL)http://dx.doi.org/10.1371/journal.pone.0021622
URIhttp://hdl.handle.net/10261/47798
DOI10.1371/journal.pone.0021622
E-ISSN1932-6203
Appears in Collections:(IBMCP) Artículos
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