Synthesis of DNA fragments in yeast by one-step assembly of overlapping oligonucleotides.
Here it is demonstrated that the yeast Saccharomyces cerevisiae can take up and assemble at least 38 overlapping single-stranded oligonucleotides and a linear double-stranded vector in one transformation event. These oligonucleotides can overlap by as few as 20 bp, and can be as long as 200 nucleotides in length. This straightforward scheme for assembling chemically-synthesized oligonucleotides could be a useful tool for building synthetic DNA molecules.
Gibson
Daniel G
DG
The J Craig Venter Institute, Synthetic Biology Group, 9704 Medical Center Drive, Rockville, MD 20850, USA. dgibson@jcvi.org
D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D009841:Oligonucleotides; D004247:DNA