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Nucleic Acids Res. 2010 May;38(8):2558-69. doi: 10.1093/nar/gkq119. Epub 2010 Mar 7.

Cloning whole bacterial genomes in yeast.

Author information

1
Synthetic Biology and Bioenergy Group, The J. Craig Venter Institute, San Diego, CA 92121, USA. gbenders@jcvi.org

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.

PMID:
20211840
PMCID:
PMC2860123
DOI:
10.1093/nar/gkq119
[Indexed for MEDLINE]
Free PMC Article

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