Synthesis of DNA fragments in yeast by one-step assembly of overlapping oligonucleotides

By D. G. Gibson. 2009

290 citations recorded by the source corpus.

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.

Subfields: Genetics, Molecular Biology

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Genetics

1. Jay D. Keasling
2. George M. Church
3. David Baker
4. Barry L. Stoddard
5. Sriram Kosuri
6. Jasmine L. Gallaher
7. Karthik Devarajan

In Molecular Biology

1. David Baker
2. Barry L. Stoddard
3. Jasmine L. Gallaher
4. George M. Church
5. Sriram Kosuri
6. A. Bar‐Even
7. Jay D. Keasling
8. Elad Noor

Citations in the corpus

• Coordinated and Compartmentalized Neuromodulation Shapes Sensory Processing in Drosophila (2015, neutral): “, 2013) (Addgene Plasmid #40753) was appended to the Drosophila synaptotagmin 1 coding sequence (DGRC Stock #4839) with an intervening 3xGS linker by PCR and Gibson Assembly (Gibson, 2009).”
• Large-scale de novo DNA synthesis: technologies and applications (2014, background): “More recently, Gibson and colleagues developed both in vivo 28, 29 and in vitro 30,31 one-step protocols for assembling and cloning oligos directly into plasmid backbones.”
• Computational protein design enables a novel one-carbon assimilation pathway (2015, methodology): “All three gene fragments were assembled into the linearized vector in one step via the method described by Gibson et al.(1).”