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Proc Natl Acad Sci U S A. 2008 Dec 23;105(51):20404-9. doi: 10.1073/pnas.0811011106. Epub 2008 Dec 10.

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

Author information

1
The J. Craig Venter Institute, Synthetic Biology Group, Rockville, MD 20850, USA. dgibson@jcvi.org

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.

PMID:
19073939
PMCID:
PMC2600582
DOI:
10.1073/pnas.0811011106
[Indexed for MEDLINE]
Free PMC Article

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