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通过大规模平行DNA测序获得个体的完整基因组。

The complete genome of an individual by massively parallel DNA sequencing.

作者信息

Wheeler David A, Srinivasan Maithreyan, Egholm Michael, Shen Yufeng, Chen Lei, McGuire Amy, He Wen, Chen Yi-Ju, Makhijani Vinod, Roth G Thomas, Gomes Xavier, Tartaro Karrie, Niazi Faheem, Turcotte Cynthia L, Irzyk Gerard P, Lupski James R, Chinault Craig, Song Xing-zhi, Liu Yue, Yuan Ye, Nazareth Lynne, Qin Xiang, Muzny Donna M, Margulies Marcel, Weinstock George M, Gibbs Richard A, Rothberg Jonathan M

机构信息

Human Genome Sequencing Center, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77030, USA.

出版信息

Nature. 2008 Apr 17;452(7189):872-6. doi: 10.1038/nature06884.

Abstract

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, 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 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'.

摘要

基因变异与疾病及药物反应之间的关联,以及核酸技术的进步,让人们对“基因组医学”的影响充满了极大的乐观情绪。然而,二倍体人类基因组规模巨大,约60亿碱基对,这使得测序方法无法常规应用于解读完整的个人人类基因组。为了实现基因组学对人类健康的全部潜力,必须克服这一限制。在此,我们报告了单一个体詹姆斯·D·沃森的二倍体基因组的DNA序列,该序列在皮升大小的反应容器中使用大规模平行测序技术,在两个月内测序达到了7.4倍冗余度。这个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个在编码序列内导致氨基酸替换。此外,我们准确鉴定了小规模(2 - 40000碱基对)的插入和缺失多态性以及拷贝数变异,这些变异导致了26000至150万个碱基对的染色体片段的大规模增减。总体而言,这些结果与近期传统方法对单一个体测序的结果非常吻合。然而,除了速度更快且成本显著降低外,这种测序技术避免了细菌克隆随机鸟枪法测序中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,我们进一步证明了获得了新的人类序列,包括传统基因组测序以前未鉴定的新基因。这是通过下一代技术测序的第一个基因组。因此,它是应对“个性化基因组测序”未来挑战的一个试点。

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