GENETIC LINKAGE MAPPING OF CHROMOSOME 13

13 号染色体的遗传连锁图谱

• 批准号: 6122438
• 负责人: ROBERT E FERRELL
• 金额: --
• 依托单位: MELLON PITTS CORPORATION (MPC CORP)
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6122438
• 关键词: biological products; biomedical resource; biotechnology; genome; human tissue

This project will construct a high resolution genetic map of human chromosome with an average resolution of 1-2 centimorgans. To achieve this goal, we will identify simple sequence repeat containing clones from chromosome 13 enriched plasmid and cosmid libraries by hybridization to poly (dA-dC)/poly (dT-dG) and other simple sequence probes. These (CA)n containing clones will be sequenced to obtained unique flanking sequences, primers for amplification by the PCR method will be selected, and each marker will be tested for length polymorphism. We will ultimately identify 100-150 di-, tri-, and tetranucleotide repeat polymorphisms with a PIC of 0.7 or greater. We will roughly assign physical location of each marker by hybridization to a YAC library and by und in situ hybridization to human metaphase chromosomes. Approximately located markers will be placed on the genetic map by linkage analysis in the extended C.E.P.H. panel. C.E.P.H. families will be typed by PCR amplification of polymorphic segments and resolution of alleles on standard sequencing gels followed by autoradiography. Use of multiplex PCR and simultaneousx analysis of multiple reactions will be developed to speed genotyping. Automated genotyping using the ABI automated sequencing system will be explored. Gene-centromere distances for markers flanking the centromere will be estimated using a panel of Type II human ovarian teratomas.

该项目将构建一张高分辨率的人类基因图谱 染色体的平均分辨率为1-2厘米器官。要实现 为了达到这个目标，我们将识别包含简单序列重复的克隆 来自13号染色体丰富的质粒库和粘粒文库 与聚(dA-DC)/聚(dt-dg)等简单序列的杂交 探测器。这些含有(CA)n的克隆将被测序以获得 独特的侧翼序列，可用于通过聚合酶链式反应方法进行扩增 将被选中，并且将测试每个标记的长度 多态。我们最终将确定100-150个二-、三-和 PIC等于或大于0.7的四核苷酸重复序列多态。我们 将通过杂交粗略地确定每个标记的物理位置 与YAC文库连接，与人中期染色体进行原位杂交 染色体。位于大致位置的标记将放置在 在扩展的C.E.P.H.小组中进行连锁分析的遗传图谱。 C.E.P.H.家系将通过聚合酶链式反应扩增多态进行分型 标准测序凝胶上等位基因的片段和拆分 然后是放射自显影。多重聚合酶链式反应和同步扩增技术的应用 将开发多反应分析，以加快基因分型。 使用ABI自动测序系统的自动基因分型将是 探索过了。侧翼标记的基因着丝粒距离 着丝粒将使用一组II型人卵巢进行评估 畸胎瘤。