Accurate multiplex gene synthesis from programmable DNA microchips.

Published

Journal Article

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, 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.

Full Text

Cited Authors

  • Tian, J; Gong, H; Sheng, N; Zhou, X; Gulari, E; Gao, X; Church, G

Published Date

  • December 2004

Published In

Volume / Issue

  • 432 / 7020

Start / End Page

  • 1050 - 1054

PubMed ID

  • 15616567

Pubmed Central ID

  • 15616567

Electronic International Standard Serial Number (EISSN)

  • 1476-4687

International Standard Serial Number (ISSN)

  • 0028-0836

Digital Object Identifier (DOI)

  • 10.1038/nature03151

Language

  • eng