| | |  | | | | Record from Web of Science® | | | | | | |
| Accurate multiplex gene synthesis from programmable DNA microchips |
|
|
| Author(s): Tian JD, Gong H, Sheng NJ, Zhou XC, Gulari E, Gao XL, Church G |
| Source: NATURE Volume: 432 Issue: 7020 Pages: 1050-1054 Published: DEC 23 2004 |
| Times Cited: 112 References: 21 |
| Abstract: Testing the many hypotheses from genomics and systems biology experiments demands accurate and cost-effective gene and genome synthesis. Here we describe a microchip-based technology for multiplex gene synthesis. Pools of thousands of 'construction' oligonucleotides and tagged complementary 'selection' oligonucleotides are synthesized on photo-programmable microfluidic chips(1), released, amplified and selected by hybridization to reduce synthesis errors ninefold. A one-step polymerase assembly multiplexing reaction assembles these into multiple genes. This technology enabled us to synthesize all 21 genes that encode the proteins of the Escherichia coli 30S ribosomal subunit, and to optimize their translation efficiency in vitro through alteration of codon bias. This is a significant step towards the synthesis of ribosomes in vitro and should have utility for synthetic biology in general. |
| Document Type: Article |
| Language: English |
| Reprint Address: Church, G (reprint author), Harvard Univ, Sch Med, 77 Ave Louis Pasteur, Boston, MA 02115 USA |
Addresses:
1. Harvard Univ, Sch Med, Boston, MA 02115 USA
2. Univ Houston, Dept Chem, Houston, TX 77004 USA
3. Atact Technol Inc, Houston, TX 77054 USA
4. Univ Michigan, Dept Chem Engn, Ann Arbor, MI 48109 USA |
| Publisher: NATURE PUBLISHING GROUP, MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND |
| Subject Category: Multidisciplinary Sciences |
| IDS Number: 881RL |
| ISSN: 0028-0836 |
| DOI: 10.1038/nature03151 |
|
| | | | | | | | | Record from Web of Science® | | | | | | | | | |