ANALYSIS AND CLONING OF A DEVELOPMENTAL SKIN LOCUS

有人建议通过基因和分子技术来分析， 一种人类基因，其表达是正常所必需的 神经外胚层发育。在这方面进行的研究 提案将针对一个人的分析 发育基因，色素性失禁(IP)。IP是X链接的 显性的半合子致死性疾病。IP携带者女性为 可辨认并具有特征的神经外胚层 发育异常。IP基因座已初步确定 已本地化到Xp11.21。 该建议提供了通过连锁对IP基因座进行分析 利用采集的多代人基因组DNA样本进行分析 IP家族和特征人类X染色体探针能够 检测限制性片段长度多态(RFLP‘s)。 包含以下内容的体细胞杂交的集合 重排的X染色体将用于分离和定位克隆 能够检测RFLP的X条染色体片段，到IP 区域。 我们建议通过鉴定来从分子水平上描绘IP基因座 与IP相关的重排X染色体的断裂点 表型。构建含IP的体细胞杂交种 X/常染色体易位将用于鉴定X染色体 IP之间的重叠、删除或断点异构性 Southern杂交分析易位与脉冲场梯度 电泳酶切图谱技术。X染色体DNA 所描绘的Ip基因座的片段将被克隆。独特的， IP基因座的高度保守区域将提供DNA 用于IP基因分离的候选片段。 这些分析将提供信息，以进一步描绘 一个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。 对神经外胚层发育的更透彻的理解，人类 发育过程、基因和基因产物 正常完成发育过程，突变 扰乱这些过程应该是这些研究的结果。

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.