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FRAMEWORK LINKAGE MAP FOR CHROMOSOME 14

14 号染色体的框架连锁图

基本信息

批准号：
2208851
负责人：
DEBORAH A NICKERSON
金额：
$ 20.67万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-04-01 至 1995-03-31
项目状态：
已结题

项目摘要

We will explore the feasibility of a genetic mapping system using polymorphic sequence-tagged sites (pSTSs) containing single nucleotide substitutions. We have developed an automated system for typing these simple biallelic markers that has several advantages over existing DNA typing methods. It is automated and has high throughput (1200 reactions/day) . It has a simple readout that is read directly by a computer. The reagents, oligonucleotides, are stable and safe (non-isotopic). The assay is easy to learn and relies on two widely used procedures, the polymerase chain reaction (PCR) and enzyme-linked immunosorbent assays (ELISAs). Furthermore , we find that clusters of linked biallelic pSTSs can be obtained with high heterozygosity (> 70%). Therefore, linked biallelic pSTSs can be as informative as manually typed simple or complex repeats. We will use biallelic pSTSs to construct a framework map of human chromosome 14 with an average marker spacing of 10 to 15 cM. We will obtain random pSTSs from genomic DNA samples using sequences from M13 libraries of flow-sorted human chromosome 14. These pSTSs will be rapidly and automatically ordered by genetic linkage analysis of informative CEPH families. Once a framework chromosome map is obtained, we will isolate YAC clones for each framework markers. This will produce physical anchors for the framework map and will also provide a source of sequences to obtain other pSTSs linked to each of the framework markers. Combined the linked biallelic pSTSs for each framework location will form an index marker (3 to 4 linked pSTSs) of high heterozygosity (> 70%). This will generate a framework map for human chromosome 14 that can be rapidly and automatically typed to detect genetic linkage of human disease-causing genes or phenotypic traits.

我们将探索使用基因作图系统的可行性含有单核苷酸的多态序列标记位点(PSTs) 替换。我们已经开发了一个自动打字系统来打字与现有DNA相比具有几个优势的简单双等位基因标记打字方法。它是自动化的，具有高吞吐量(1200 反应/天)。它有一个简单的读数，可以由一个电脑。这种试剂，寡核苷酸，是稳定和安全的 (非同位素)。该分析方法简单易学，并依赖于两个广泛使用的程序、聚合酶链式反应(PCR)和酶联反应免疫吸附试验(ELISA)。此外，我们还发现，连锁双等位基因pSTS杂合度高(&gt；70%)。因此，链接的双等位基因pSTS可以像手动键入的那样提供信息简单或复杂的重复。我们将利用双等位基因pSTSS构建人类的框架图 14号染色体，平均标记间距为10~15 cM。我们会利用M13的序列从基因组DNA样本中获得随机的pSTS 流动排序的人类14号染色体文库。这些pSTS将迅速并通过信息性CEPH的遗传连锁分析自动排序家人。一旦获得框架染色体图谱，我们将分离YAC 每个框架标记的克隆。这将为以下各项产生物理锚该框架图还将提供序列来源以获得链接到每个框架标记的其他pSTS。组合了链接的每个骨架位置的双等位pSTS将形成索引标记(3 到4个连锁的高杂合度(&gt；70%)的pSTS。这将生成一个人类14号染色体的框架图，可以快速和自动地用来检测人类致病基因的遗传连锁或表型性状。