Shotgun sequencing of the human genome
Shotgun sequencing of the human genome
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DOI：
10.1126/science.280.5369.1540
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发表时间：
1998-06-05
期刊：
影响因子：
56.9
通讯作者：
Hunkapiller, M
中科院分区：
文献类型：
--
作者：
Venter, JC;Adams, MD;Hunkapiller, M
The Human Genome Project (HGP) was officially launched in the United States on 1 October 1990 as a 15-year program to map and sequence the complete set of human chromosomes and those of several model organisms. The HGP is laying the groundwork for a revolution in medicine and biology. Its importance is underscored by the level of funding from the National Institutes of Health, the Department of Energy (DOE), the Wellcome Trust, and other governments and foundations around the world.From the inception of the HGP, major technical innovations that would affect its timetable and cost were considered essential to success. The development of bacterial artificial chromosomes (BACs)(1) provided a key advance. BACs are propagated in Escherichia coli and carry large [∼ 150-kilobase pairs (kbp)] inserts stably. In contrast, ordered cosmid clones that served as the basis of yeast (2) and Caenorhabditis elegans (3) genome sequencing projects are less stable and much shorter (∼ 35 kbp). Fluorescent labeling of DNA fragments generated by the Sanger dideoxy chain termination method has been the mainstay of almost all large-scale sequencing projects since the introduction of the first semi-automated sequencer by Applied Biosystems in 1987 and the development of Taq cycle sequencing in 1990. New models of the sequencer that can process more samples, Taq polymerase engineered especially for sequencing, and higher sensitivity dyes have improved throughput, accuracy, and operating costs. Publication of the first genome from a self-replicating organism, Haemophilus influenzae, was based on a whole-genome shotgun (random sequencing) method (4). A set of algorithms called the TIGR Assembler (5) together with scaffolding sequences from both ends of 18-kbp inserts in bacteriophage lambda clones were critical for determination of correct order and assembly. Eight additional genomes have since been completed by these methods (4, 6, 7), and several others are nearing completion, including genomes with high GC (∼ 65%) and high AT (∼ 82%) composition, which present special problems for sequencing and assembly.
