COLLAGEN GENES IN HERITABLE CONNECTIVE TISSUE DISORDERS

我的总体目标是既接受教学训练，又接受实践训练。 要成为一名独立的生物医学工作者，必须有实验室经验 调查员。我计划将这一培训应用于研究贡献 各种遗传性疾病的胶原基因改变， 包括选定的纤维结缔组织遗传性疾病和 软骨营养不良症。现建议采取以下方法： (1)各种胶原基因的结构和组织 紊乱将通过Southern印迹分析来确定。不寻常的图案 将与在控件面板中看到的控件进行比较，以确定 前者是严重的基因改变的结果。 (2)胶原基因的结构将发生更微妙的变化 通过连锁分析检测到。多个受影响的大家庭 将对个体进行“突变”表型的共分离检查。 和一个特定的胶原等位基因，其遗传通过多态检测 限制点。 (3)(1)或(2)表示突变的个人或家庭 在胶原基因中，对该基因编码的信使核糖核酸和蛋白质的研究将 以研究该基因的功能意义 改变，突变基因将被克隆和测序以证明 突变的本质。 (4)染色体定位未知的胶原蛋白基因座 或未经证实，将承担此类任务。 (5)不同胶原基因间的遗传距离 (1)和(2)和选定的共线基因座将被确定。 这些研究的目的有四个：(一)确定和 描述胶原基因突变的特征，这些基因与 结缔组织的某些家族性疾病，(Ii)确定 这些突变对胶原mRNA和蛋白质合成的影响 结构；(Iii)识别结构中的正常变化和 限制性内切酶片段揭示胶原基因的组织 长度多态；(Iv)为了增加我们对人类基因图谱的了解， 在染色体定位和遗传距离方面 在紧密连锁的基因座和各种胶原基因之间。

My overall goal is to acquire both the didactic training and practical laboratory experience necesary to become an independent biomedical investigator. I plan to apply this training to studies of the contribution of collagen gene alterations to a variety of hereditary disorders, including selected heritable disorders of fibrous connective tissue and chondrodystrophies. The following approaches are proposed: (1) The structure and organization of the various collagen genes in these disorders will be determined by Southern blot analysis. Unusual patterns will be compared to those seen in a panel of controls to determine if the former are the result of gross gene alterations. (2) More subtle alterations in the structure of collagen genes will be detected by linkage analysis. Large families with multiple affected individuals will be examined for cosegregation of the "mutant" phenotype and a specific collagen allele whose inheritance is detected by polymorphic restriction sites. (3) In individuals or families in whom either (1) or (2) indicates mutation in a collagen gene, studies of mRNA and protein encoded by this gene will be performed to investigate the functional significance of the gene alteration, and the mutant gene will be cloned and sequenced to demonstrate the nature of the mutation. (4) For those collagen loci for which the chromosomal assignment is unknown or unconfirmed, such assignments will be undertaken. (5) The genetic distance between the various collagen genes studies in (1)\and (2) and selected syntenic loci will be determined. The purpose of these studies is four-fold: (i) to identify and characterize mutations in the collagen genes which are responsible for certain familial disorders of connective tissues, (ii) to determine the effect of such mutations on collagen mRNA and protein synthesis and structure; (iii) to identify normal variations in the structure and organization of the collagen genes as revealed by restriction fragment length polymorphisms; (iv) to add to our knowledge of the human gene map, both in terms of the chromosomal localization of and the genetic distances between closely linked loci and the various collagen genes.