One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome

Daniel G. Gibson; Gwynedd A. Benders; Kevin C. Axelrod; Jayshree Zaveri; Mikkel A. Algire; Monzia Moodie; Michael G. Montague; J. Craig Venter; Hamilton O. Smith; Clyde A. Hutchison

doi:10.1073/pnas.0811011106

One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome

Daniel G. Gibson, Gwynedd A. Benders, Kevin C. Axelrod, Jayshree Zaveri, Mikkel A. Algire, Monzia Moodie, Michael G. Montague, J. Craig Venter, Hamilton O. Smith, Clyde A. Hutchison

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

306 Scopus citations

Abstract

We previously reported assembly and cloning of the synthetic Mycoplasma genitalium JCVI-1.0 genome in the yeast Saccharomyces cerevisiae by recombination of six overlapping DNA fragments to produce a 592-kb circle. Here we extend this approach by demonstrating assembly of the synthetic genome from 25 overlapping fragments in a single step. The use of yeast recombination greatly simplifies the assembly of large DNA molecules from both synthetic and natural fragments.

Original language	English (US)
Pages (from-to)	20404-20409
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	105
Issue number	51
DOIs	https://doi.org/10.1073/pnas.0811011106
State	Published - Dec 23 2008
Externally published	Yes

Keywords

Combinatorial assembly
Genome synthesis
In vivo DNA assembly
Mycoplasma genitalium
Synthetic biology
Yeast transformation

ASJC Scopus subject areas

General

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10.1073/pnas.0811011106

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Gibson, D. G., Benders, G. A., Axelrod, K. C., Zaveri, J., Algire, M. A., Moodie, M., Montague, M. G., Venter, J. C., Smith, H. O., & Hutchison, C. A. (2008). One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome. Proceedings of the National Academy of Sciences of the United States of America, 105(51), 20404-20409. https://doi.org/10.1073/pnas.0811011106

One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome. / Gibson, Daniel G.; Benders, Gwynedd A.; Axelrod, Kevin C. et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, No. 51, 23.12.2008, p. 20404-20409.

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

Gibson, DG, Benders, GA, Axelrod, KC, Zaveri, J, Algire, MA, Moodie, M, Montague, MG, Venter, JC, Smith, HO & Hutchison, CA 2008, 'One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome', Proceedings of the National Academy of Sciences of the United States of America, vol. 105, no. 51, pp. 20404-20409. https://doi.org/10.1073/pnas.0811011106

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AU - Montague, Michael G.

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One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome

Abstract

Keywords

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