Daniel G. Gibson1, Gwynedd A. Benders2, Kevin Axelrod3, Jayshree Zaveri3, Mikkel A. Algire3, Monzia Moodie3, Michael Montague3, J. Craig Venter3, Hamilton O. Smith2, Clyde A. Hutchison2
1The J. Craig Venter Institute, Synthetic Biology Group, Rockville, MD 20850, USA.
2The J. Craig Venter Institute, Synthetic Biology Group, San Diego, CA 92121
3The J. Craig Venter Institute, Synthetic Biology Group, Rockville, MD 20850 and
Tóm tắt
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.
