PORCINE ENAMEL PROTEINS

描述：（改编自研究者摘要）长期 目的是确定牙釉质的分子机制， 形成和鉴定负责釉质形成的基因 （大赦国际）。 提出了四个具体目标：1）克隆和鉴定cDNA 编码猪釉蛋白，29 kDa的钙结合蛋白，釉原蛋白， 2）克隆猪釉原蛋白基因并鉴定其结构 基因，确定猪釉原蛋白基因的数量，以及它们的 染色体定位; 3）克隆和表征人cDNA， 编码釉蛋白和29 kDa钙结合蛋白的基因组克隆 (CBP)并确定其染色体定位; 4）测试 釉基质蛋白基因与釉质形成的连锁关系 你好 猪釉基质蛋白氨基端序列分析 是由合作者Makato Mingae博士提供的。 mRNA编码 这些蛋白质被克隆并通过克隆PCR充分表征 扩增产物，5'和3' cDNA末端快速扩增（RACE）， 并通过直接筛选猪成釉器官上皮特异性λ cDNA文库 猪釉原蛋白基因分离自雄性 猪基因组文库。 基因的结构是由 PCR扩增产物的表征、限制性分析，以及 DNA测序 釉原蛋白基因的数量由Southern杂交确定。 印迹分析 染色体定位是通过荧光标记来确定的。 原位杂交 人cDNA和基因组克隆分离自 人牙齿特异性cDNA文库和λ基因组文库。 的 染色体定位是通过荧光原位 杂交方法 连锁分析是在显示 显性遗传的人工智能使用候选基因方法。 特定突变 通过单链构象多态性分析鉴定， 其特征在于DNA测序。

DESCRIPTION: (Adapted from investigator's Abstract) The long term objectives are to determine the molecular mechanisms of dental enamel formation and to identify the genes responsible for amelogenesis imperfecta (AI). Four specific aims are proposed: 1) to clone and characterize cDNAs encoding porcine enamelin, the 29 kDa calcium binding protein, amelogenin, tuftelin and sheathlin; 2) to clone and characterize the porcine amelogenin gene(s), determine the number of porcine amelogenin gene(s), and their chromosomal localization(s); 3) to clone and characterize human cDNA and genomic clones encoding enamelin and the 29 kDa calcium binding protein (CBP) and to determine their chromosomal localizations; 4) to test for linkage between genes encoding enamel matrix proteins and amelogenesis imperfecta. The amino-terminal sequence of porcine enamel matrix proteins has been provided by a collaborator, Dr. Makato Fukae. The mRNA encoding these proteins are cloned and fully characterized by cloning PCR amplification products, 5' and 3' rapid amplification of cDNA ends (RACE), and by direct screening of a porcine enamel organ epithelia-specific Lambda cDNA library. The porcine amelogenin gene(s) is isolated from a male porcine genomic library. The structure of the gene is determined by the characterization of PCR amplification products, restriction analyses, and DNA sequencing. The number of amelogenin genes is determined by Southern blot analysis. The chromosomal localization is determined by fluorescent in situ hybridization. The human cDNA and genomic clones are isolated from a human tooth-specific cDNA library and a Lambda genomic library. The chromosomal localizations are determined by fluorescent in situ hybridization. Linkage analysis is performed on pedigrees displaying dominantly inherited AI using a candidate gene approach. Specific mutations are identified by single stranded conformatinal polymorphism analysis and characterized by DNA sequencing.