One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome
Top Cited Papers
- 23 December 2008
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 105 (51) , 20404-20409
- https://doi.org/10.1073/pnas.0811011106
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
This publication has 18 references indexed in Scilit:
- Complete Chemical Synthesis, Assembly, and Cloning of a Mycoplasma genitalium GenomeScience, 2008
- Rebuilding microbial genomesBioEssays, 2007
- Direct cloning of full-length mouse mitochondrial DNA using a Bacillus subtilis genome vectorGene, 2007
- Nonhomologous End Joining in YeastAnnual Review of Genetics, 2005
- Combining two genomes in one cell: Stable cloning of the Synechocystis PCC6803 genome in the Bacillus subtilis 168 genomeProceedings of the National Academy of Sciences, 2005
- Rapid generation of long synthetic tandem repeats and its application for analysis in human artificial chromosome formationNucleic Acids Research, 2005
- Transfer of YACs up to 2.3 Mb intact into human cells with polyethylenimineGene Therapy, 1999
- General Method for Plasmid Construction Using Homologous RecombinationBioTechniques, 1999
- Eukaryotic replicators and associated protein complexesCurrent Opinion in Genetics & Development, 1995
- A Model System to Assess the Integrity of Mammalian YACs during Transformation and Propagation in YeastGenomics, 1994