ANALYSIS AND CLONING OF A DEVELOPMENTAL SKIN LOCUS

建议通过遗传和分子技术， 一种人类基因，其表达对于正常的 神经外胚层发育 在此进行的研究 建议将针对人类的分析 发育基因，色素失禁症（IP）。 IP为X连锁 显性半合子致死性疾病 IP携带者女性 可识别且具有特征性神经外胚层 发育异常 IP轨迹已初步 定位于Xp11.21。 该建议提供了通过连锁分析IP基因座 分析使用收集的基因组DNA样品从多代 IP家族和表征的人类X染色体探针能够 限制性片段长度多态性（RFLP）的检测。 体细胞杂种的集合，包含 重排的X染色体将用于分离和定位克隆 能够检测RFLP的X染色体片段， 地区 我们建议从分子水平上描绘IP基因座， 与IP相关的重排X染色体的断点 表型 含有IP的构建体细胞杂种 X/常染色体易位将用于鉴定X染色体 IP之间的重叠、缺失或断点异质性 通过Southern印迹分析和脉冲场梯度检测易位 电泳限制性作图技术。 X染色体DNA 将克隆来自描绘的Ip基因座的片段。 独一无二， IP基因座的高度保守区域将提供DNA 用于分离IP基因的候选片段。 这些分析将提供信息，以进一步描述 人类发育基因的遗传和染色体定位 为IP基因的克隆提供了必要的起点。 更深入地了解神经外胚层发育，人类 发育过程、基因和基因产物， 发育过程的正常完成，以及 应该从这些研究中得出结论。

A proposal is made to analyze, by genetic and molecular techniques, a human gene whose expression is essential for normal neuroectodermal development. The research undertaken in this proposal will be directed towards the analysis of a human developmental gene, incontinentia pigmenti (IP). IP is an X-linked dominant, hemizygote lethal disorder. IP carrier females are identifiable and present with characteristic neuroectodermal developmental abnormalities. The IP locus has been tentatively localized to Xp11.21. This proposal provides for analysis of the IP locus by linkage analysis using collected genomic DNA samples from multigenerational IP families and characterized human X chromosomal probes capable of detecting restriction fragment length polymorphisms (RFLP's). An assemble collection of somatic cell hybrids containing rearranged X chromosomes will be used to isolated and map cloned X chromosomal fragments capable of detecting RFLP's, to the IP region. We propose to molecularly delineate the IP locus by identifying breakpoints of rearranged X chromosomes associated with the IP phenotype. Constructed somatic cell hybrids containing IP X/autosomal translocations will be used to identify X chromosomal overlaps, deletions, or breakpoint heterogeneity among the IP translocations by Southern blot analysis and pulse field gradient electrophoretic restriction mapping techniques. X chromosomal DNA fragments from the delineated Ip locus will be cloned. Unique, highly conserved regions from the IP locus will provide DNA fragment candidates for the isolation of the IP gene. These analyses will provide information to further delineate the genetic and chromosomal localization of a human developmental gene and provide the necessary starting point for cloning the IP gene. A more thorough understanding of neuroectodermal development, human developmental processes, genes and gen products necessary for normal completion of developmental process, and mutations that disrupt these processes should result from these studies.