Cloning whole bacterial genomes in yeast.
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.
Benders
Gwynedd A
GA
Synthetic Biology and Bioenergy Group, The J. Craig Venter Institute, San Diego, CA 92121, USA. gbenders@jcvi.org
Noskov
Vladimir N
VN
Denisova
Evgeniya A
EA
Lartigue
Carole
C
Gibson
Daniel G
DG
Assad-Garcia
Nacyra
N
Chuang
Ray-Yuan
RY
Carrera
William
W
Moodie
Monzia
M
Algire
Mikkel A
MA
Phan
Quang
Q
Alperovich
Nina
N
Vashee
Sanjay
S
Merryman
Chuck
C
Venter
J Craig
JC
Smith
Hamilton O
HO
Glass
John I
JI
Hutchison
Clyde A
CA
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*