title: Cloning whole bacterial genomes in yeast.
abstract: Most microbes have not been cultured, and many of those that are cultivatable are difficult, dangerous or expensive to propagate or are genetically intractable. Routine cloning of large genome fractions or whole genomes from these organisms would significantly enhance their discovery and genetic and functional characterization. Here we report the cloning of whole bacterial genomes in the yeast Saccharomyces cerevisiae as single-DNA molecules. We cloned the genomes of Mycoplasma genitalium (0.6 Mb), M. pneumoniae (0.8 Mb) and M. mycoides subspecies capri (1.1 Mb) as yeast circular centromeric plasmids. These genomes appear to be stably maintained in a host that has efficient, well-established methods for DNA manipulation.
authors.affiliation: Synthetic Biology and Bioenergy Group, The J. Craig Venter Institute, San Diego, CA 92121, USA. gbenders@jcvi.org
authors.forename: Gwynedd A
authors.initials: GA
authors.lastname: Benders
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Noskov
authors.forename: Evgeniya A
authors.initials: EA
authors.lastname: Denisova
authors.forename: Carole
authors.initials: C
authors.lastname: Lartigue
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Gibson
authors.forename: Nacyra
authors.initials: N
authors.lastname: Assad-Garcia
authors.forename: Ray-Yuan
authors.initials: RY
authors.lastname: Chuang
authors.forename: William
authors.initials: W
authors.lastname: Carrera
authors.forename: Monzia
authors.initials: M
authors.lastname: Moodie
authors.forename: Mikkel A
authors.initials: MA
authors.lastname: Algire
authors.forename: Quang
authors.initials: Q
authors.lastname: Phan
authors.forename: Nina
authors.initials: N
authors.lastname: Alperovich
authors.forename: Sanjay
authors.initials: S
authors.lastname: Vashee
authors.forename: Chuck
authors.initials: C
authors.lastname: Merryman
authors.forename: J Craig
authors.initials: JC
authors.lastname: Venter
authors.forename: Hamilton O
authors.initials: HO
authors.lastname: Smith
authors.forename: John I
authors.initials: JI
authors.lastname: Glass
authors.forename: Clyde A
authors.initials: CA
authors.lastname: Hutchison
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004171:Diploidy; D005822:Genetic Vectors / Q000737:chemistry; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics*; D045704:Mycoplasma genitalium / Q000235:genetics; D009176:Mycoplasma mycoides / Q000235:genetics; D009177:Mycoplasma pneumoniae / Q000235:genetics; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*