POLYMORPHIC AND EXPRESSED MARKERS FOR HUMAN CHROMOSOME 3

该项目的长期目标是生产一组多晶型 3号染色体的标记物，其将用作作图的框架， 测序染色体。 目前，3号染色体图谱是稀疏的， 只有少数基因被定位在这条大染色体上。 将为每条染色体分离大约250个标记，并将其放置在 在辐射杂交和连锁图谱上。 该项目的最终结果将 是一个2厘米的地图和物理标记大约每兆字节。 的 待开发的标记物将是非常有用的，因为它们是非常有用的。 多态性和基于PCR的。 该项目的第一个具体目标是 产生基于简单重复的高度多态性标记。 重复2次， 3和4碱基对将从染色体特异性文库中分离， 并且它们的序列用于开发基于PCR的测定。 第二个目标是 在定位于3号染色体的已知基因座内出现多态性。 的 单链构象多态性（SSCP）和测序方法 将鉴定多态性，该多态性将通过 等位基因特异性寡核苷酸。 将在2010年开发更多的标记物。 第三个具体目标是那些缺少标记的染色体区域。 ALU PCR将用于辐射减少的杂交体，以产生区域 特定克隆 将对这些克隆进行测序和分析， 多态 所有上述标记将被放置在 辐射杂种的物理图谱和CEPH分析的遗传图谱， 其他家族DNA 因为这个项目将产生基于PCR的多态性， 将立即对科学界有用。 调查人员 寻找连锁标记将有一个巨大的水库， 标记来绘制。 我们的方法吸收了所有已知的数据， 3号染色体，将已知的序列纳入详细的物理和 基因图谱 这些标记是定向比对的第一步 的YAC和粘粒克隆，并将促进关闭在这条染色体上， 未来

The long term goal of this project to produce a battery of polymorphic markers for chromosome 3 which will serve as a framework for mapping and sequencing the chromosome. Currently, the chromosome 3 maps is sparse and only a handful of gene have been mapped to this large chromosome. Approximately 250 markers will be isolated for each chromosome and placed on radiation hybrid and linkage maps. The net result of this project will be a 2 cM map and physical markers approximately every megabase. The markers to be developed will be highly useful as they are highly polymorphic and PCR based. The first specific aim of the project is to produce highly polymorphic markers base on simple repeats. Repeats of 2, 3, and 4 base pairs will be isolated from chromosome specific libraries, and their sequence used to develop a PCR based assay. A second aim is to develop polymorphisms within known gene loci mapped to chromosome 3. The method of single strand conformational polymorphism (SSCP) and sequencing will identify polymorphism which will be detected in individuals by an allele specific oligonucleotide. Further markers will be developed in specific aim three for those regions of the chromosomes which lack markers. ALU PCR will be used on radiation reduced hybrids to generate region specific clones. These clones will be sequenced and analyzed for polymorphism. All the above mentioned markers will be placed on the physical map by radiation hybrids and the genetic map by analyzing CEPH and other family DNA. Because this project will generate PCR based polymorphisms, the markers will be immediately useful to the scientific community. Investigators seeking linkage markers will have a large reservoir of precisely mapped markers to draw upon. Our approach assimilates all the known data on chromosome 3 by incorporating known sequences into detailed physical and genetic maps. These markers are a first step towards a directed aligning of YAC and cosmid clones and will facilitate closure on this chromosome in the future.