CHARACTERIZATION OF MAPPED HUMAN BAC CLONES

细菌人工染色体(BAC)克隆在从基因组测序到基因发现的广泛项目中使用。我们的实验室正在鉴定和组装一组绘制的人类BAC克隆。希望使用这些资源的调查人员可以使用这些资源。我们资源中的BAC克隆被锚定在辐射杂交图谱中的序列标记位点(STS)标记上。克隆的地图信息可以在我们的基于网络的数据库GenMapDB上免费获得。随着我们映射克隆以添加到这个资源中，很明显，如果我们能够进一步描述克隆的特征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。在本申请中，我们建议通过实现以下目标来实现这些目标。1.在所有作图克隆上制备DNA，用于进一步分析，包括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鉴定(目标2)。2.通过进行HindIII指纹分析和末端测序进一步鉴定克隆。3.升级现有数据库，并使用FISH分析数据评估我们基于RH的BAC图的准确性。有关克隆的更多信息将提供一种基因组试剂，可用于染色体突变图谱、基因图谱和多态筛选等项目。在鉴定克隆的过程中，我们还将获得信息，使我们深入了解RH图、基因组序列图和细胞遗传学图之间的符合率。

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.