The complete genome of an individual by massively parallel DNA sequencing
The complete genome of an individual by massively parallel DNA sequencing
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DOI：
10.1038/nature06884
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发表时间：
2008-04-17
期刊：
影响因子：
64.8
通讯作者：
Rothberg, Jonathan M.
中科院分区：
文献类型：
--
作者：
Wheeler, David A.;Srinivasan, Maithreyan;Rothberg, Jonathan M.
遗传变异与疾病和药物反应的关联，以及核酸技术的改进，给“基因组医学”的影响带来了极大的乐观。然而，人类二倍体基因组的惊人大小(1)，大约6千兆碱基，阻碍了测序方法的常规应用来破译完整的人类个体基因组。为了充分发挥基因组学对人类健康的潜力，必须克服这一限制。在这里，我们报告了单个个体James D. Watson的二倍体基因组的DNA序列，在两个月内使用皮升大小的反应容器进行了大规模平行测序，获得了7.4倍的冗余。该序列在两个月内完成，成本约为传统毛细管电泳方法的百分之一。将该序列与参考基因组进行比较，鉴定出330万个单核苷酸多态性，其中10654个导致编码序列内的氨基酸取代。此外，我们准确地鉴定了小规模（2 - 40000个碱基对）的插入和删除多态性以及拷贝数变异，导致染色体片段的大规模增加和减少，范围从26000到150万个碱基对。总的来说，这些结果与最近用传统方法对单个个体(2)进行测序的结果一致。然而，除了更快和显著更便宜之外，这种测序技术避免了细菌克隆随机鸟枪测序固有的基因组序列的任意丢失，因为它在无细胞系统中扩增DNA。因此，我们进一步展示了新的人类序列的获取，包括以前未被传统基因组测序鉴定的新基因。这是第一个基因组测序的下一代技术。因此，它是未来挑战“个性化基因组测序”的一个试点。
The association of genetic variation with disease and drug response, and improvements in nucleic acid technologies, have given great optimism for the impact of 'genomic medicine'. However, the formidable size of the diploid human genome(1), approximately 6 gigabases, has prevented the routine application of sequencing methods to deciphering complete individual human genomes. To realize the full potential of genomics for human health, this limitation must be overcome. Here we report the DNA sequence of a diploid genome of a single individual, James D. Watson, sequenced to 7.4- fold redundancy in two months using massively parallel sequencing in picolitre- size reaction vessels. This sequence was completed in two months at approximately one-hundredth of the cost of traditional capillary electrophoresis methods. Comparison of the sequence to the reference genome led to the identification of 3.3 million single nucleotide polymorphisms, of which 10,654 cause amino- acid substitution within the coding sequence. In addition, we accurately identified small-scale ( 2 - 40,000 base pair ( bp)) insertion and deletion polymorphism as well as copy number variation resulting in the large- scale gain and loss of chromosomal segments ranging from 26,000 to 1.5 million base pairs. Overall, these results agree well with recent results of sequencing of a single individual(2) by traditional methods. However, in addition to being faster and significantly less expensive, this sequencing technology avoids the arbitrary loss of genomic sequences inherent in random shotgun sequencing by bacterial cloning because it amplifies DNA in a cell- free system. As a result, we further demonstrate the acquisition of novel human sequence, including novel genes not previously identified by traditional genomic sequencing. This is the first genome sequenced by next- generation technologies. Therefore it is a pilot for the future challenges of 'personalized genome sequencing'.