One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome

Daniel G. Gibson, Gwynedd A. Benders, Kevin C. Axelrod, Jayshree Zaveri, Mikkel A. Algire, Monzia Moodie, Michael Montague, J. Craig Venter, Hamilton O. Smith, Clyde A. Hutchison

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)20404-20409
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume105
Issue number51
DOIs
StatePublished - Dec 23 2008
Externally publishedYes

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Keywords

  • Combinatorial assembly
  • Genome synthesis
  • In vivo DNA assembly
  • Mycoplasma genitalium
  • Synthetic biology
  • Yeast transformation

ASJC Scopus subject areas

  • General

Cite this

Gibson, D. G., Benders, G. A., Axelrod, K. C., Zaveri, J., Algire, M. A., Moodie, M., Montague, M., Venter, J. C., Smith, H. O., & Hutchison, C. A. (2008). One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome. Proceedings of the National Academy of Sciences of the United States of America, 105(51), 20404-20409. https://doi.org/10.1073/pnas.0811011106