One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome
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.
- Publication:
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Proceedings of the National Academy of Science
- Pub Date:
- December 2008
- DOI:
- 10.1073/pnas.0811011106
- Bibcode:
- 2008PNAS..10520404G
- Keywords:
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- Biological Sciences:Genetics