GoldenBraid: an iterative simple standardized cloning system system for standardized assembly assembly of recyclable reusable genetic modules
| Autor: | Sarrión Perdigones, Manuel Alejandro, Juárez Ortega, Paloma, Fernández Del Carmen, María Asunción, Granell Richart, Antonio, Orzáez Calatayud, Diego Vicente |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: | |
| Zdroj: | RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia instname |
| Popis: | Synthetic 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. This work was supported by the Spanish Ministry of Science and Innovation grants BIO2008-03434 and BIO2010-15384. A. Sarrion-Perdigones is a recipient of a FPI fellowship of the Spanish Ministry of Science and Innovation and P. Juarez is a recipient of a FPU fellowship from the Spanish Ministry of Education. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. |
| Databáze: | OpenAIRE |
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