title: Broad-host-range vector system for synthetic biology and biotechnology in cyanobacteria.
abstract: Inspired by the developments of synthetic biology and the need for improved genetic tools to exploit cyanobacteria for the production of renewable bioproducts, we developed a versatile platform for the construction of broad-host-range vector systems. This platform includes the following features: (i) an efficient assembly strategy in which modules released from 3 to 4 donor plasmids or produced by polymerase chain reaction are assembled by isothermal assembly guided by short GC-rich overlap sequences. (ii) A growing library of molecular devices categorized in three major groups: (a) replication and chromosomal integration; (b) antibiotic resistance; (c) functional modules. These modules can be assembled in different combinations to construct a variety of autonomously replicating plasmids and suicide plasmids for gene knockout and knockin. (iii) A web service, the CYANO-VECTOR assembly portal, which was built to organize the various modules, facilitate the in silico construction of plasmids, and encourage the use of this system. This work also resulted in the construction of an improved broad-host-range replicon derived from RSF1010, which replicates in several phylogenetically distinct strains including a new experimental model strain Synechocystis sp. WHSyn, and the characterization of nine antibiotic cassettes, four reporter genes, four promoters, and a ribozyme-based insulator in several diverse cyanobacterial strains.
authors: Division of Biological Sciences, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA aataton@gmail.com.
authors: Arnaud
authors: A
authors: Taton
authors: Division of Biological Sciences, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
authors: Federico
authors: F
authors: Unglaub
authors: Division of Biological Sciences, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
authors: Nicole E
authors: NE
authors: Wright
authors: Division of Biological Sciences, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
authors: Wei Yue
authors: WY
authors: Zeng
authors: Scripps Institution of Oceanography, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA Institut de Biologie de I'Ecole Normale Supérieure, CNRS, UMR 8197, 46 rue d'Ulm, 75230 Paris, France.
authors: Javier
authors: J
authors: Paz-Yepes
authors: Scripps Institution of Oceanography, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
authors: Bianca
authors: B
authors: Brahamsha
authors: Scripps Institution of Oceanography, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
authors: Brian
authors: B
authors: Palenik
authors: Synthetic Biology Division, Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008, USA.
authors: Todd C
authors: TC
authors: Peterson
authors: Synthetic Biology Division, Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008, USA.
authors: Farzad
authors: F
authors: Haerizadeh
authors: Division of Biological Sciences, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
authors: Susan S
authors: SS
authors: Golden
authors: Division of Biological Sciences, University of California San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA.
authors: James W
authors: JW
authors: Golden
mesh_terms: D001709:Biotechnology / Q000379:methods; D003198:Computer Simulation; D000458:Cyanobacteria / Q000235:genetics*; D004352:Drug Resistance, Microbial / Q000235:genetics; D018390:Gene Targeting; D017930:Genes, Reporter; D005822:Genetic Vectors; D010957:Plasmids / Q000235:genetics*; D012093:Replicon; D058615:Synthetic Biology / Q000379:methods