title: Creating bacterial strains from genomes that have been cloned and engineered in yeast.
abstract: We recently reported the chemical synthesis, assembly, and cloning of a bacterial genome in yeast. To produce a synthetic cell, the genome must be transferred from yeast to a receptive cytoplasm. Here we describe methods to accomplish this. We cloned a Mycoplasma mycoides genome as a yeast centromeric plasmid and then transplanted it into Mycoplasma capricolum to produce a viable M. mycoides cell. While in yeast, the genome was altered by using yeast genetic systems and then transplanted to produce a new strain of M. mycoides. These methods allow the construction of strains that could not be produced with genetic tools available for this bacterium.
authors.affiliation: J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850, USA.
authors.forename: Carole
authors.initials: C
authors.lastname: Lartigue
authors.forename: Sanjay
authors.initials: S
authors.lastname: Vashee
authors.forename: Mikkel A
authors.initials: MA
authors.lastname: Algire
authors.forename: Ray-Yuan
authors.initials: RY
authors.lastname: Chuang
authors.forename: Gwynedd A
authors.initials: GA
authors.lastname: Benders
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Noskov
authors.forename: Evgeniya A
authors.initials: EA
authors.lastname: Denisova
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Gibson
authors.forename: Nacyra
authors.initials: N
authors.lastname: Assad-Garcia
authors.forename: Nina
authors.initials: N
authors.lastname: Alperovich
authors.forename: David W
authors.initials: DW
authors.lastname: Thomas
authors.forename: Chuck
authors.initials: C
authors.lastname: Merryman
authors.forename: Clyde A
authors.initials: CA
authors.lastname: Hutchison
authors.forename: Hamilton O
authors.initials: HO
authors.lastname: Smith
authors.forename: J Craig
authors.initials: JC
authors.lastname: Venter
authors.forename: John I
authors.initials: JI
authors.lastname: Glass
mesh_terms: D002503:Centromere; D003001:Cloning, Molecular; D019175:DNA Methylation; D004262:DNA Restriction Enzymes / Q000235:genetics / Q000378:metabolism; D015263:Deoxyribonucleases, Type III Site-Specific / Q000235:genetics; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D016680:Genome, Bacterial; D045623:Mycoplasma capricolum / Q000235:genetics*; D009176:Mycoplasma mycoides / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014169:Transformation, Bacterial
chemical_list: D004262:DNA Restriction Enzymes; D015263:Deoxyribonucleases, Type III Site-Specific