Bouffard G G, Idol J R, Braden V V, Iyer L M, Cunningham A F, Weintraub L A, Touchman J W, Mohr-Tidwell R M, Peluso D C, Fulton R S, Ueltzen M S, Weissenbach J, Magness C L, Green E D
Genome Technology Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Genome Res. 1997 Jul;7(7):673-92. doi: 10.1101/gr.7.7.673.
The construction of highly integrated and annotated physical maps of human chromosomes represents a critical goal of the ongoing Human Genome Project. Our laboratory has focused on developing a physical map of human chromosome 7, a approximately 170-Mb segment of DNA that corresponds to an estimated 5% of the human genome. Using a yeast artificial chromosome (YAC)-based sequence-tagged site (STS)-content mapping strategy, 2150 chromosome 7-specific STSs have been established and mapped to a collection of YACs highly enriched for chromosome 7 DNA. The STSs correspond to sequences generated from a variety of DNA sources, with particular emphasis placed on YAC insert ends, genetic markers, and genes. The YACs include a set of relatively nonchimeric clones from a human-hamster hybrid cell line as well as clones isolated from total genomic libraries. For map integration, we have localized 260 STSs corresponding to Genethon genetic markers and 259 STSs corresponding to markers orders by radiation hybrid (RH) mapping on our YAC contigs. Analysis of the data with the program SEGMAP results in the assembly of 22 contigs that are "anchored" on the Genethon genetic map, the RH map, and/or the cytogenetic map. These 22 contigs are ordered relative to one another, are (in all but 3 cases) oriented relative to the centromere and telomeres, and contain > 98% of the mapped STSs. The largest anchored YAC contig, accounting for most of 7p, contains 634 STSs and 1260 YACs. An additional 14 contigs, accounting for approximately 1.5% of the mapped STSs, are assembled but remain unanchored on either the genetic or RH map. Therefore, these 14 "orphan" contigs are not ordered relative to other contigs. In our contig maps, adjacent STSs are connected by two or more YACs in > 95% of cases. With 2150 mapped STSs, our map provides an average STS spacing of approximately 79 kb. The physical map we report here exceeds the goal of 100-kb average STS spacing and should provide an excellent framework for systematic sequencing of the chromosome.
构建高度整合且带有注释的人类染色体物理图谱是正在进行的人类基因组计划的一个关键目标。我们实验室专注于绘制人类7号染色体的物理图谱,这是一段约170兆碱基对的DNA片段,约占人类基因组的5%。利用基于酵母人工染色体(YAC)的序列标签位点(STS)含量作图策略,已建立了2150个7号染色体特异性STS,并将其定位到一组高度富集7号染色体DNA的YAC文库中。这些STS对应于从多种DNA来源产生的序列,特别着重于YAC插入末端、遗传标记和基因。这些YAC包括一组来自人 - 仓鼠杂交细胞系的相对非嵌合克隆以及从全基因组文库中分离的克隆。为了进行图谱整合,我们已将对应于Genethon遗传标记的260个STS和对应于辐射杂种(RH)作图标记顺序的259个STS定位到我们的YAC重叠群上。用SEGMAP程序分析数据后,组装出22个重叠群,这些重叠群“锚定”在Genethon遗传图谱、RH图谱和/或细胞遗传图谱上。这22个重叠群相互之间是有序排列的,(除3种情况外)相对于着丝粒和端粒是定向的,并且包含了超过98%的已定位STS。最大的锚定YAC重叠群占7号染色体短臂(7p)的大部分,包含634个STS和1260个YAC。另外还有14个重叠群被组装起来,但在遗传图谱或RH图谱上仍未锚定,它们占已定位STS的约1.5%。因此,这14个“孤立”重叠群与其他重叠群之间没有顺序关系。在我们的重叠群图谱中,超过95%的情况下相邻的STS由两个或更多YAC连接。有了2150个已定位的STS,我们的图谱平均STS间距约为79千碱基对。我们在此报告的物理图谱超过了平均STS间距100千碱基对的目标,应为该染色体的系统测序提供一个出色的框架。