CHARACTERIZATION OF MAPPED HUMAN BAC CLONES

细菌人工染色体（BAC）克隆广泛应用于从基因组测序到基因发现的各种项目中。我们实验室正在鉴定和组装一组人类BAC克隆图谱。此资源可供希望使用它们的调查人员使用。我们的资源中的BAC克隆锚定在辐射杂交图谱中的序列标记位点（STS）标记上。克隆的映射信息可以在我们基于web的数据库GenMapDB上免费获得。当我们将克隆添加到这个资源中时，很明显，如果我们能够进一步表征克隆并确保分配给每个克隆的基因组位置是正确的，那么这个集合的价值将大大增加。在本应用程序中，我们建议通过实现以下目标来实现这些目标。1. 为所有绘制的克隆准备DNA，以便进一步分析，包括由我们的合作者进行的荧光原位杂交（FISH）和我们自己的进一步表征（目的2）。通过进行HindIII指纹图谱和末端测序进一步表征克隆。3. 升级现有数据库，并利用FISH分析数据评估基于rh的BAC地图的准确性。有关克隆的额外信息将提供一种基因组试剂，可用于诸如染色体突变图谱、基因图谱和多态性筛选等项目。在对克隆进行表征的过程中，我们还将获得一些信息，让我们了解RH图、基因组序列图和细胞遗传学图之间的一致性率。

Bacterial artificial chromosome (BAC) clones are used in a wide range of projects from genome sequencing to gene discovery. Our laboratory is identifying and assembling a set of mapped human BAC clones. This resource is available for investigators who wish to use them. The BAC clones in our resource are anchored to sequence tagged site (STS) markers in radiation hybrid maps. The mapping information of the clones is freely available on our web-based database, GenMapDB. As we mapped clones to add to this resource, it became clear that the value of this collection would be greatly increased if we could further characterize the clones and ensure that the genomic location assigned to each clone is correct. In this application, we propose to achieve these goals by accomplishing the following aims. 1. To prepare DNA on all the mapped clones for further analysis including fluorescent in situ hybridization (FISH) carried out by our collaborators and our own further characterization (Aim 2). 2. To further characterize the clones by performing HindIII fingerprinting and end sequencing. 3. To upgrade the current database and to assess the accuracy of our RH-based BAC map with data from FISH analysis. The additional information about the clones will provide a genomic reagent that can be used in projects such as mapping of chromosomal mutations, gene mapping and polymorphism screening. In the process of characterizing the clones, we will also get information that gives us insight into the concordance rate between RH map, genome sequence map and cytogenetic map.