1. Cloning whole bacterial genomes in yeast
- Author
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Hamilton O. Smith, Monzia Moodie, Mikkel A. Algire, Daniel G. Gibson, Sanjay Vashee, Quang Phan, Nacyra Assad-Garcia, John I. Glass, Clyde A. Hutchison, Ray-Yuan Chuang, Gwynedd A. Benders, Carole Lartigue, J. Craig Venter, Chuck Merryman, Vladimir N. Noskov, Nina Alperovich, Evgeniya A. Denisova, William Carrera, and The J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, Maryland 20850, USA
- Subjects
[SDV]Life Sciences [q-bio] ,Saccharomyces cerevisiae ,Genetic Vectors ,Molecular Sequence Data ,Mycoplasma genitalium ,Bacterial genome size ,Genome ,03 medical and health sciences ,chemistry.chemical_compound ,Plasmid ,Mycoplasma ,Genetics ,Cloning, Molecular ,ComputingMilieux_MISCELLANEOUS ,030304 developmental biology ,Cloning ,Recombination, Genetic ,0303 health sciences ,biology ,Base Sequence ,030306 microbiology ,Mycoplasma mycoides ,biology.organism_classification ,Diploidy ,Yeast ,3. Good health ,Mycoplasma pneumoniae ,chemistry ,Synthetic Biology and Chemistry ,DNA ,Genome, Bacterial - 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.
- Published
- 2010
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