GENETIC LINKAGE MAPPING OF CHROMOSOME 13

13 号染色体的遗传连锁图谱

• 批准号: 6282473
• 负责人: ROBERT E FERRELL
• 金额: $ 1.19万
• 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/6282473
• 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.

该项目将构建人类高分辨率基因图谱 染色体平均分辨率为1-2厘摩。 达到 为了这个目标，我们将识别包含克隆的简单序列重复 来自 13 号染色体富集的质粒和粘粒文库 与poly (dA-dC)/poly (dT-dG) 和其他简单序列杂交 探针。 这些含有 (CA)n 的克隆将被测序以获得 独特的侧翼序列、PCR 方法扩增引物 将被选择，并且将测试每个标记的长度 多态性。 我们最终将确定 100-150 个二、三和 PIC 为 0.7 或更高的四核苷酸重复多态性。 我们 将通过杂交粗略地分配每个标记的物理位置 至 YAC 文库并通过原位杂交至人中期 染色体。 大致定位的标记将放置在 通过扩展 C.E.P.H 中连锁分析的遗传图谱控制板。 C.E.P.H.通过PCR扩增多态性对家族进行分型 标准测序凝胶上等位基因的片段和分辨率 随后进行放射自显影。 使用多重 PCR 和同步 x 将开发多重反应分析以加速基因分型。 使用 ABI 自动测序系统进行自动基因分型将 探索过。 侧翼标记的基因着丝粒距离 将使用一组 II 型人类卵巢来估计着丝粒 畸胎瘤。