Accurate multiplex gene synthesis from programmable DNA microchips

被引：295

作者：

Tian, JD

Gong, H

Sheng, NJ

Zhou, XC

Gulari, E

Gao, XL

Church, G

机构：

[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

来源：

NATURE | 2004年 / 432卷 / 7020期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1038/nature03151

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：1050 / 1054

页数：5

共 21 条

[1] PREDICTING DNA DUPLEX STABILITY FROM THE BASE SEQUENCE [J].