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...

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Detalles Bibliográficos
Autores principales: Benders, Gwynedd A., Noskov, Vladimir N., Denisova, Evgeniya A., Lartigue, Carole, Gibson, Daniel G., Assad-Garcia, Nacyra, Chuang, Ray-Yuan, Carrera, William, Moodie, Monzia, Algire, Mikkel A., Phan, Quang, Alperovich, Nina, Vashee, Sanjay, Merryman, Chuck, Venter, J. Craig, Smith, Hamilton O., Glass, John I., Hutchison, Clyde A.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2010
Materias:
Synthetic Biology and Chemistry
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2860123/
https://www.ncbi.nlm.nih.gov/pubmed/20211840
http://dx.doi.org/10.1093/nar/gkq119
Descripción
Sumario: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.

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