title: One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome.
abstract: We previously reported assembly and cloning of the synthetic Mycoplasma genitalium JCVI-1.0 genome in the yeast Saccharomyces cerevisiae by recombination of six overlapping DNA fragments to produce a 592-kb circle. Here we extend this approach by demonstrating assembly of the synthetic genome from 25 overlapping fragments in a single step. The use of yeast recombination greatly simplifies the assembly of large DNA molecules from both synthetic and natural fragments.
authors.affiliation: The J. Craig Venter Institute, Synthetic Biology Group, Rockville, MD 20850, USA. dgibson@jcvi.org
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Gibson
authors.forename: Gwynedd A
authors.initials: GA
authors.lastname: Benders
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Axelrod
authors.forename: Jayshree
authors.initials: J
authors.lastname: Zaveri
authors.forename: Mikkel A
authors.initials: MA
authors.lastname: Algire
authors.forename: Monzia
authors.initials: M
authors.lastname: Moodie
authors.forename: Michael G
authors.initials: MG
authors.lastname: Montague
authors.forename: J Craig
authors.initials: JC
authors.lastname: Venter
authors.forename: Hamilton O
authors.initials: HO
authors.lastname: Smith
authors.forename: Clyde A
authors.initials: CA
authors.lastname: Hutchison
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000096:biosynthesis*; D016680:Genome, Bacterial / Q000235:genetics*; D045704:Mycoplasma genitalium / Q000235:genetics*; D009838:Oligodeoxyribonucleotides / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D015003:Yeasts / Q000235:genetics*
chemical_list: D009838:Oligodeoxyribonucleotides; D004247:DNA