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; Zaveri, Jayshree; Algire, Mikkel A; Moodie, Monzia; Montague, Michael G; Venter, J Craig et al. · Proc Natl Acad Sci U S A · 2008

basic_science · Level V

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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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