END SEQUENCING AND FINGERPRINTING OF MOUSE BAC LIBRARIES

随着人类基因组测序项目转向生产测序，一项针对小鼠基因组测序的计划正在制定中。目前的计划采用“序列第一，地图第二”的策略，与人类计划一样，该策略基于对BAC克隆的鸟枪测序，该克隆包括选定小鼠菌株（C57BL6/J）基因组的连续区域。以细菌人工染色体（BAC）为载体构建的文库因其克隆能力强、稳定性好而成为高通量基因组测序的首选。目前的全基因组方法已在人类基因组测序计划中得到验证，该方法使用BAC末端序列和限制性指纹图谱来构建BAC contigs。限制性内切指纹图谱不仅可以作为基于BAC端序列组装的BAC contigs的独立确认，还可以用于鉴定嵌合BAC和内部缺失的克隆。这种随用随测策略在构造序列就绪映射时节省了大量的时间和精力。本提案是在TIGR进行BAC端测序，在Clemson University Genomics Institute Physical Mapping Center进行限制性内切指纹鉴定，使用小鼠BAC文库或为支持小鼠基因组测序而构建的文库。克隆、序列、指纹凝胶图像以及使用FPC程序（C. Soderlund， Sanger中心）对克隆进行分类，将为那些对小鼠基因组进行测序的人以及整个社区提供可用的资源。我们将继续关注BAC克隆端测序和限制性基因指纹项目的质量，同时开发和实施自动化和新方法，以降低成本和提高产量。

As the human project shifts into production sequencing, a plan is developing for sequencing the mouse genome. The current plan employs a "sequence first, map second" strategy which, like the human project, is based on shotgun sequencing of BAC clones comprising contiguous regions of the genome of the selected mouse strain (C57BL6/J). Libraries constructed in bacterial artificial chromosomes (BAC) vectors have become the choice for high throughput genomic sequencing projects because of their cloning capacity and high stability. The current whole-genome approach, which has been validated in the human genome sequencing project, uses BAC end sequences and restriction fingerprints to build BAC contigs. Restriction fingerprints not only serve as independent confirmation of BAC contigs assembled based on BAC end sequence, but also enable identification of chimeric BACs and clones with internal deletions. This map-as-you-go strategy saves substantial time and effort in constructing sequence ready maps. This proposal is to perform BAC end sequencing at TIGR and restriction fingerprinting at the Clemson University Genomics Institute Physical Mapping Center from the mouse BAC library or libraries constructed to support mouse genome sequencing. The clones, the sequences, gel images of the fingerprints, and the binning of the clones using the FPC program (C. Soderlund, Sanger Centre) will be an available resource for those sequencing the mouse genome, and the community at large. We will continue our focus on quality in this BAC clone end sequencing and restriction fingerprinting project while developing and implementing automation and new methodologies for reducing costs and increasing throughput.