Creating bacterial strains from genomes that have been cloned and engineered in yeast.
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.
Lartigue
Carole
C
J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850, USA.
Vashee
Sanjay
S
Algire
Mikkel A
MA
Chuang
Ray-Yuan
RY
Benders
Gwynedd A
GA
Ma
Li
L
Noskov
Vladimir N
VN
Denisova
Evgeniya A
EA
Gibson
Daniel G
DG
Assad-Garcia
Nacyra
N
Alperovich
Nina
N
Thomas
David W
DW
Merryman
Chuck
C
Hutchison
Clyde A
CA
Smith
Hamilton O
HO
Venter
J Craig
JC
Glass
John I
JI
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
D004262:DNA Restriction Enzymes; D015263:Deoxyribonucleases, Type III Site-Specific