Enzymatic assembly of DNA molecules up to several hundred kilobases

被引：4
|
作者：
Gibson, Daniel G. [1 ]
Young, Lei [1 ]
Chuang, Ray-Yuan [1 ]
Venter, J. Craig [1 ]
Hutchison, Clyde A., III
Smith, Hamilton O.
机构：
[1] J Craig Venter Inst, Synthet Biol Grp, Rockville, MD USA
来源：
NATURE METHODS | 2009年 / 6卷 / 05期
基金：
美国能源部;
关键词：
ESCHERICHIA-COLI; POLYMERASE; CLONING; RECOMBINATION; EXONUCLEASE; PCR;
D O I：
10.1038/NMETH.1318
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
We describe an isothermal, single-reaction method for assembling multiple overlapping DNA molecules by the concerted action of a 5' exonuclease, a DNA polymerase and a DNA ligase. First we recessed DNA fragments, yielding single-stranded DNA overhangs that specifically annealed, and then covalently joined them. This assembly method can be used to seamlessly construct synthetic and natural genes, genetic pathways and entire genomes, and could be a useful molecular engineering tool.
引用
收藏
页码：343 / U41
页数：5
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