High-resolution BAC-based map of the central portion of mouse chromosome 5.
High-resolution BAC-based map of the central portion of mouse chromosome 5.
复制标题
小鼠 5 号染色体中央部分基于 BAC 的高分辨率图谱。
DOI：
10.1101/gr.195101
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发表时间：
2001
期刊：
影响因子：
7
通讯作者：
Bucan,M
中科院分区：
文献类型：
--
作者：
Crabtree,J;Wiltshire,T;Brunk,B;Zhao,S;Schug,J;StoeckertJr,CJ;Bucan,M
目前的小鼠基因组测序策略涉及全基因组鸟枪法与基于克隆的测序相结合。高分辨率的物理图将为组装连续的序列片段提供基础。我们已经建立了一个基于细菌人工染色体（BAC）的小鼠5号染色体5-Mb区域的图谱，包括三个基因家族：受体酪氨酸激酶（PdgfraKit-Kdr），非受体蛋白酪氨酸型激酶（Tec-Txk）和神经递质GABA的A型受体（Gabra 2，Gabrb 1，Gabrg 1和Gabra 4）。通过使用已知基因和序列标记位点（STS）杂交筛选C57 BL/6 J（RPCI-23）BAC文库来启动BAC重叠群的构建。通过搜索RPCI-23和RPCI-24文库的可用限制性指纹的数据库来鉴定另外的重叠克隆。这一努力导致选择了>600个BAC克隆、251 kb的BAC末端序列，以及在该5-Mb区域内放置了40个已知和/或预测的基因。我们使用这个高分辨率的地图来说明BAC指纹图谱与辐射混合地图通过组装表达序列标签（EST）的整合。从三个代表性的BAC克隆的注释，我们证明，高达98%的草案序列的每个重叠群可以使用已知的基因，BAC结束，转录组件的共识序列，并与orthopathic人类序列的比较，排序和定向。对于功能研究，当序列片段组装成50-200-kb的片段时对其进行注释将是非常有价值的。
The current strategy for sequencing the mouse genome involves the combination of a whole-genome shotgun approach with clone-based sequencing. High-resolution physical maps will provide a foundation for assembling contiguous segments of sequence. We have established a bacterial artificial chromosome (BAC)-based map of a 5-Mb region on mouse Chromosome 5, encompassing three gene families: receptor tyrosine kinases (PdgfraKit-Kdr), nonreceptor protein-tyrosine type kinases (Tec–Txk), and type-A receptors for the neurotransmitter GABA (Gabra2, Gabrb1, Gabrg1, andGabra4). The construction of a BAC contig was initiated by hybridization screening the C57BL/6J (RPCI-23) BAC library, using known genes and sequence tagged sites (STSs). Additional overlapping clones were identified by searching the database of available restriction fingerprints for the RPCI-23 and RPCI-24 libraries. This effort resulted in the selection of >600 BAC clones, 251 kb of BAC-end sequences, and the placement of 40 known and/or predicted genes within this 5-Mb region. We use this high-resolution map to illustrate the integration of the BAC fingerprint map with a radiation-hybrid map via assembled expressed sequence tags (ESTs). From annotation of three representative BAC clones we demonstrate that up to 98% of the draft sequence for each contig could be ordered and oriented using known genes, BAC ends, consensus sequences for transcript assemblies, and comparisons with orthologous human sequence. For functional studies, annotation of sequence fragments as they are assembled into 50–200-kb stretches will be remarkably valuable.