MOLECULAR MODELING OF DOPAMINE ANTAGONISTS

该项目将构建一个高分辨率的人类基因图谱， 染色体，平均分辨率为1-2厘摩。 实现 为了达到这一目标，我们将鉴定含有简单重复序列的克隆， 从13号染色体富集的质粒和粘粒文库， 与聚（dA-dC）/聚（dT-dG）和其它简单序列杂交 probes. 将对这些含有（CA）2的克隆进行测序以获得 独特的侧翼序列，通过PCR方法扩增的引物 将被选中，每个标记将被测试长度 多态 我们将最终确定100-150个二，三， 四核苷酸重复多态性，PIC为0.7或更高。 我们 将通过杂交粗略地分配每个标记的物理位置 YAC文库，并通过与人中期分裂相的原位杂交 染色体 大致定位的标记将放置在 在扩展的C.E.P.H.中通过连锁分析绘制遗传图谱。面板 CEPH将通过PCR扩增多态性位点对家族进行分型。 标准测序凝胶上等位基因的片段和分辨率 然后进行放射自显影。 使用多重PCR和PCR扩增 将开发对多种反应的分析，以加快基因分型。 将使用ABI自动测序系统进行自动基因分型。 探讨了 基因-着丝粒距离的侧翼标记的 着丝粒将使用一组II型人卵巢癌细胞来估计。 畸胎瘤

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)2 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 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 amplifica tion 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.