Letters to Nature

Nature 432, 1050-1054 (23 December 2004) | doi:10.1038/nature03151; Received 26 April 2004; Accepted 22 October 2004

Accurate multiplex gene synthesis from programmable DNA microchips

Jingdong Tian1, Hui Gong1, Nijing Sheng2, Xiaochuan Zhou3, Erdogan Gulari4, Xiaolian Gao2 & George Church1

  1. Harvard Medical School, 77 Ave Louis Pasteur, Boston, Massachusetts 02115, USA
  2. Department of Chemistry, University of Houston, Houston, Texas 77004-5003, USA
  3. Atactic Technologies Inc., 2575 West Bellfort, Suite 270, Houston, Texas 77054, USA
  4. Chemical Engineering Department, University of Michigan, Ann Arbor, Michigan 48109-2136, USA

Correspondence to: George Church1 Email: g1m1c1@arep.med.harvard.edu
The 14.6-kb operon of 21 ribosomal genes has been deposited in GenBank under accession number AY773199.

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 chips1, 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.

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