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

The J. Craig Venter Institute, Synthetic Biology Group, Rockville, MD 20850, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.81). 01/2009; 105(51):20404-9. DOI: 10.1073/pnas.0811011106
Source: PubMed

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.

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