Gene synthesis by circular assembly amplification

被引：53

作者：

Bang, Duhee ^{[1
]}

Church, George M. ^{[1
]}

机构：

[1] Harvard Univ, Sch Med, Dept Genet, Boston, MA 02115 USA

来源：

NATURE METHODS | 2008年 / 5卷 / 01期

关键词：

D O I：

10.1038/NMETH1136

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Here we report the development of a gene-synthesis technology, circular assembly amplification. In this approach, we first constructed exonuclease-resistant circular DNA via simultaneous ligation of oligonucleotides. Exonuclease- and subsequent mismatch cleaving endonuclease-mediated degradation of the resulting ligation mixture eliminated error-rich products, thereby substantially improving gene-synthesis quality. We used this method to construct genes encoding a small thermostable DNA polymerase, a highly repetitive DNA sequence and large (>4 kb) constructs.

引用

页码：37 / 39

页数：3

共 14 条

[1] Protein-mediated error correction for de novo DNA synthesis -: art. no. e162
Carr, PA
Park, JS
Lee, YJ
Yu, T
Zhang, SG
Jacobson, JM
[J]. NUCLEIC ACIDS RESEARCH, 2004, 32 (20) : e162
[2] Removal of mismatched bases from synthetic genes by enzymatic mismatch cleavage
Fuhrmann, M
Oertel, W
Berthold, P
Hegemann, P
[J]. NUCLEIC ACIDS RESEARCH, 2005, 33 (06) : 1 - 8
[3] USER fusion: a rapid and efficient method for simultaneous fusion and cloning of multiple PCR products
Geu-Flores, Fernando
Nour-Eldin, Hussam H.
Nielsen, Morten T.
Halkier, Barbara A.
[J]. NUCLEIC ACIDS RESEARCH, 2007, 35 (07)
[4] A highly active synthetic mammalian retrotransposon
Han, JS
Boeke, JD
[J]. NATURE, 2004, 429 (6989) : 314 - 318
[5] Total synthesis of long DNA sequences: Synthesis of a contiguous 32-kb polyketide synthase gene cluster
Kodumal, SJ
Patel, KG
Reid, R
Menzella, HG
Welch, M
Santi, DV
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (44) : 15573 - 15578
[6] Crystal structure of a Y-family DNA polymerase in action: A mechanism for error-prone and lesion-bypass replication
Ling, H
Boudsocq, F
Woodgate, R
Yang, W
[J]. CELL, 2001, 107 (01) : 91 - 102
[7] Combinatorial polyketide biosynthesis by de novo design and rearrangement of modular polyketide synthase genes
Menzella, HG
Reid, R
Carney, JR
Chandran, SS
Reisinger, SJ
Patel, KG
Hopwood, DA
Santi, DV
[J]. NATURE BIOTECHNOLOGY, 2005, 23 (09) : 1171 - 1176
[8] Richardson SM, 2006, GENOME RES, V16, P550, DOI 10.1101/gr.4431306
[9] A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics
SantaLucia, J
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (04) : 1460 - 1465
[10] Generating a synthetic genome by whole genome assembly:: φX174 bacteriophage from synthetic oligonucleotides
Smith, HO
Hutchison, CA
Pfannkoch, C
Venter, JC
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (26) : 15440 - 15445

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