Accurate multiplex gene synthesis from programmable DNA microchips

Jingdong Tian, Hui Gong, Nijing Sheng, Xiaochuan Zhou, Erdogan Gulari, Xiaolian Gao and George Church ()
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Jingdong Tian: Harvard Medical School
Hui Gong: Harvard Medical School
Nijing Sheng: University of Houston
Xiaochuan Zhou: Atactic Technologies Inc.
Erdogan Gulari: University of Michigan
Xiaolian Gao: University of Houston
George Church: Harvard Medical School

Nature, 2004, vol. 432, issue 7020, 1050-1054

Abstract: 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 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.

Date: 2004
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DOI: 10.1038/nature03151

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