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One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome |
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| Authors: | Daniel G. Gibson Gwynedd A. Benders Kevin C. Axelrod Jayshree Zaveri Mikkel A. Algire Monzia Moodie Michael G. Montague J. Craig Venter Hamilton O. Smith Clyde A. Hutchison III |
| Affiliation: | aThe J. Craig Venter Institute, Synthetic Biology Group, Rockville, MD 20850 and ;bThe J. Craig Venter Institute, Synthetic Biology Group, San Diego, CA 92121 |
| 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. |
| Keywords: | in vivo DNA assembly genome synthesis combinatorial assembly yeast transformation Mycoplasma genitalium synthetic biology |