Gene Synthesis Method Based on Overlap Extension PCR and DNAWorks Program
Gene Synthesis Method Based on Overlap Extension PCR and DNAWorks Program
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基于重叠延伸PCR和DNAWorks程序的基因合成方法
DOI：
10.1007/978-1-62703-625-2_2
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发表时间：
2013-01-01
期刊：
影响因子：
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通讯作者：
Zhang, Li-Ping
中科院分区：
文献类型：
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作者：
Li, Gang;Dong, Bing-Xue;Zhang, Li-Ping
在后基因组时代，利用化学方法合成基因为修饰基因和探索基因的结构、表达和功能提供了强有力的工具。然而，最近基因合成技术的瓶颈是寡核苷酸合成和合成后测序的高成本。在这里，我们描述了一种基于重叠延伸PCR （OE-PCR）和DNAWorks程序的简单、快速、低成本的基因合成方法。该方法可以合成200 bp到3 kb的DNA序列，而且错误很少，而且这些错误可以很容易地通过定点突变来纠正。因此，它适用于不同基因的多路合成自动化，具有高通量基因合成的潜力。
Gene synthesis by chemical methods provides a powerful tool for modifying genes and exploring their structure, expression, and function in the post-genomic era. However, a bottleneck in recent gene synthesis technologies is the high cost of oligonucleotide synthesis and post-synthesis sequencing. Here, we describe a simple, rapid, and low-cost gene synthesis method based on overlap extension PCR (OE-PCR) and the DNAWorks program. This method enables DNA sequences with sizes ranging from 200 bp to 3 kb to be synthesized with few errors, and these errors can be easily corrected by site-directed mutagenesis. Thus, it is amenable to automation for the multiplexed synthesis of different genes and has a potential for high-throughput gene synthesis.