DNA LINKAGE STUDIES OF DEGENERATIVE RETINAL DISEASES

这项建议是为了继续我们的DNA连锁研究， 患有退行性视网膜疾病的家族，例如常染色体 显性视网膜色素变性（ADRP）和Usher综合征。 的 目标是找到这些紧密连锁的DNA标记， 疾病，将疾病位点定位到特定的染色体区域 并利用这些连锁的DNA标记来表征疾病 的位点 1986年，国家视网膜色素变性基金会成立， 一个合作项目，以促进DNA连锁研究， 视网膜色素变性（RP）。 Daiger博士是项目协调员， 这个戒酒会这 该方案的目的是确定和 表征适当的RP家族，以发送家族血液样本 到人类遗传突变细胞库进行制备， 储存转化的细胞系，并提供这些细胞系 送到戴格尔博士的实验室进行准备，测试和处理 的DNA。 目前，RP收集包含来自以下的91个细胞系： 一个大的ADRP家族和77个来自扩展Usher的品系 综合征家族 集合应该增加50到100行 1988年 我们已经准备并测试了大部分 样本，并已获得91个额外的DNA从其他RP 家庭 该提案是针对该计划的研究部分， DNA连锁检测和数据分析 除了有 我们和其他人已经在这些 我们已经测试了22种以上的DNA探针， 家庭 我们建议用20种新的方法测试150到200个人， 探针每年使用传统的Southern凝胶分析。 我们将 还开发和应用了一种新的，高效的程序， 检测人类DNA中的“mini-VNTR”多态性， 使用聚合酶链式反应（PCR）方法扩增。 目前有数百种多态性DNA探针 我们假设这些数字 将找到的微型VNTR。 方法：组织培养、基因组DNA提取、制备 DNA探针Southern凝胶分析、DNA多态性检测 通过PCR扩增，两点和多点连接 分析. 当链接建立后，我们将测试其他 家庭，并将使用分子方法来改善信息 标记的内容，开发更接近的标记，并建立 侧翼标记 ADRP和Usher综合征的连锁标志物 在早期诊断、检测遗传性 异质性，并最终，在隔离和表征 突变基因

This proposal is for funds to continue our DNA linkage studies of families with degenerative retinal diseases such as autosomal dominant retinitis pigmentosa (ADRP) and Usher's syndrome. The objectives are to find tightly linked DNA markers for these diseases, to map the disease loci to specific chromosomal regions and to use these linked DNA markers in characterizing the disease loci. In 1986 the National Retinitis Pigmentosa Foundation established a collaborative program to facilitate DNA linkage studies of retinitis pigmentosa (RP). Dr. Daiger is Program Coordinator for this program. The aims of the program are to identify and characterize appropriate RP families, to send family blood samples to the Human Genetic Mutant Cell Repository for preparation and storage of transformed cell lines, and to provide these cell lines to Dr. Daiger's laboratory for preparation, testing and handling of DNAs. At present the RP collection contains 91 cell lines from one large ADRP family and 77 lines from an extended Usher's syndrome family. The collection should increase by 50 to 100 lines in 1988. We have prepared and tested DNAs from most of these samples and have acquired 91 additional DNAs from other RP families. This proposal is for the research component of the program, that is, DNA linkage testing and data analysis. In addition to the classical genetic markers we and others have tested in these families, we already have tested over 22 DNA probes in selected families. We propose to test 150 to 200 individuals with 20 new probes per year using conventional Southern gel analysis. We will also develop and apply a novel, high efficiency procedure for detecting "mini-VNTR" polymorphisms in human DNA, based on DNA amplification using the polymerase chain reaction (pcr) method. There are hundreds of polymorphic DNA probes currently available from the scientific community and we hypothesize that like numbers of mini VNTRs will be found. Methods are tissue culture, extraction of genomic DNAs, preparation of probe DNAs Southern gel analysis, detection of DNA polymorphisms by pcr amplification, and two point and multipoint linkage analysis. When linkage is established we will test additional families and will use molecular methods to improve the information content of the marker, develop closer markers and establish flanking markers. Linked markers for ADRP and Usher's syndrome will be of value in early diagnosis, in detection of genetic heterogeneity and eventually, in isolation and characterization of the mutant genes.