Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, United Kingdom.
PLoS One. 2010 Feb 8;5(2):e9089. doi: 10.1371/journal.pone.0009089.
Physical and linkage mapping underpin efforts to sequence and characterize the genomes of eukaryotic organisms by providing a skeleton framework for whole genome assembly. Hitherto, linkage and physical "contig" maps were generated independently prior to merging. Here, we develop a new and easy method, BAC HAPPY MAPPING (BAP mapping), that utilizes BAC library pools as a HAPPY mapping panel together with an Mbp-sized DNA panel to integrate the linkage and physical mapping efforts into one pipeline. Using Arabidopsis thaliana as an exemplar, a set of 40 Sequence Tagged Site (STS) markers spanning approximately 10% of chromosome 4 were simultaneously assembled onto a BAP map compiled using both a series of BAC pools each comprising 0.7x genome coverage and dilute (0.7x genome) samples of sheared genomic DNA. The resultant BAP map overcomes the need for polymorphic loci to separate genetic loci by recombination and allows physical mapping in segments of suppressed recombination that are difficult to analyze using traditional mapping techniques. Even virtual "BAC-HAPPY-mapping" to convert BAC landing data into BAC linkage contigs is possible.
物理图谱和连锁图谱为真核生物基因组测序和特征分析提供了骨架框架,是推动这一工作的基础。到目前为止,连锁图谱和物理“连续图谱”是在合并之前独立生成的。在这里,我们开发了一种新的简单方法 BAC HAPPY MAPPING(BAP 作图),它利用 BAC 文库池作为 HAPPY 作图面板,结合 Mbp 大小的 DNA 面板,将连锁作图和物理作图的工作整合到一个流程中。以拟南芥为模式生物,我们构建了一个 BAP 图谱,该图谱整合了一系列包含 0.7x 基因组覆盖度的 BAC 池和稀释(0.7x 基因组)的基因组 DNA 片段,图谱上同时包含了大约 10%的染色体 4 的一组 40 个序列标签位点(STS)标记。所得的 BAP 图谱克服了需要通过重组分离遗传位点的多态性位点的需要,并允许在使用传统作图技术难以分析的抑制重组的片段中进行物理作图。甚至可以通过虚拟“BAC-HAPPY 作图”将 BAC 定位数据转换为 BAC 连锁连续图谱。