One-step Assembly In Yeast of 25 Overlapping DNA Fragments to Form a Complete Synthetic Mycoplasma genitalium Genome
Proc Natl Acad Sci U S A. 2008 Dec 23; 105(51): 20404-9.
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.