Cloning whole bacterial genomes in yeast

Gwynedd A. Benders; Vladimir N. Noskov; Evgeniya A. Denisova; Carole Lartigue; Daniel G. Gibson; Nacyra Assad-Garcia; Ray Yuan Chuang; William Carrera; Monzia Moodie; Mikkel A. Algire; Quang Phan; Nina Alperovich; Sanjay Vashee; Chuck Merryman; J. Craig Venter; Hamilton O. Smith; John I. Glass; Clyde A. Hutchison

doi:10.1093/nar/gkq119

Cloning whole bacterial genomes in yeast

Gwynedd A. Benders
, Vladimir N. Noskov
, Evgeniya A. Denisova
, Carole Lartigue
, Daniel G. Gibson
, Nacyra Assad-Garcia
, Ray Yuan Chuang
, William Carrera
, Monzia Moodie
, Mikkel A. Algire
, Quang Phan
, Nina Alperovich
, Sanjay Vashee
, Chuck Merryman
, J. Craig Venter
, Hamilton O. Smith
, John I. Glass
, Clyde A. Hutchison

Research output: Contribution to journal › Article › peer-review

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.

Original language	English (US)
Article number	gkq119
Pages (from-to)	2558-2569
Number of pages	12
Journal	Nucleic acids research
Volume	38
Issue number	8
DOIs	https://doi.org/10.1093/nar/gkq119
State	Published - Mar 6 2010
Externally published	Yes

ASJC Scopus subject areas

Genetics

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10.1093/nar/gkq119

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Benders, G. A., Noskov, V. N., Denisova, E. A., Lartigue, C., Gibson, D. G., Assad-Garcia, N., Chuang, R. Y., Carrera, W., Moodie, M., Algire, M. A., Phan, Q., Alperovich, N., Vashee, S., Merryman, C., Venter, J. C., Smith, H. O., Glass, J. I., & Hutchison, C. A. (2010). Cloning whole bacterial genomes in yeast. Nucleic acids research, 38(8), 2558-2569. Article gkq119. https://doi.org/10.1093/nar/gkq119

Benders, GA, Noskov, VN, Denisova, EA, Lartigue, C, Gibson, DG, Assad-Garcia, N, Chuang, RY, Carrera, W, Moodie, M, Algire, MA, Phan, Q, Alperovich, N, Vashee, S, Merryman, C, Venter, JC, Smith, HO, Glass, JI & Hutchison, CA 2010, 'Cloning whole bacterial genomes in yeast', Nucleic acids research, vol. 38, no. 8, gkq119, pp. 2558-2569. https://doi.org/10.1093/nar/gkq119

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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.",

author = "Benders, \{Gwynedd A.\} and Noskov, \{Vladimir N.\} and Denisova, \{Evgeniya A.\} and Carole Lartigue and Gibson, \{Daniel G.\} and Nacyra Assad-Garcia and Chuang, \{Ray Yuan\} and William Carrera and Monzia Moodie and Algire, \{Mikkel A.\} and Quang Phan and Nina Alperovich and Sanjay Vashee and Chuck Merryman and Venter, \{J. Craig\} and Smith, \{Hamilton O.\} and Glass, \{John I.\} and Hutchison, \{Clyde A.\}",

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AU - Benders, Gwynedd A.

AU - Noskov, Vladimir N.

AU - Denisova, Evgeniya A.

AU - Lartigue, Carole

AU - Gibson, Daniel G.

AU - Assad-Garcia, Nacyra

AU - Chuang, Ray Yuan

AU - Carrera, William

AU - Moodie, Monzia

AU - Algire, Mikkel A.

AU - Phan, Quang

AU - Alperovich, Nina

AU - Vashee, Sanjay

AU - Merryman, Chuck

AU - Venter, J. Craig

AU - Smith, Hamilton O.

AU - Glass, John I.

AU - Hutchison, Clyde A.

PY - 2010/3/6

Y1 - 2010/3/6

N2 - 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.

AB - 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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Cloning whole bacterial genomes in yeast

Abstract

ASJC Scopus subject areas

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