COLLAGEN GENES IN HERITABLE CONNECTIVE TISSUE DISORDERS

我的总体目标是获得教学培训和实践 成为独立生物医学所必需的实验室经验 调查员。 我计划将这次培训应用于贡献研究 胶原蛋白基因改变导致多种遗传性疾病， 包括选定的纤维结缔组织遗传性疾病和 软骨营养不良。 建议采用以下方法： (1)这些胶原蛋白基因的结构和组织 疾病将通过Southern印迹分析来确定。 不寻常的图案 将与控制面板中看到的进行比较，以确定是否 前者是基因总体改变的结果。 (2) 胶原蛋白基因结构发生更细微的改变 通过连锁分析检测。 多个受影响的大家庭 将检查个体是否存在“突变”表型的共分离 以及通过多态性检测其遗传的特定胶原等位基因 限制位点。 （3）在（1）或（2）表明突变的个人或家庭中 在胶原蛋白基因中，对该基因编码的 mRNA 和蛋白质的研究将 进行以研究基因的功能意义 改变，突变基因将被克隆和测序以证明 突变的性质。 (4) 对于那些染色体分配未知的胶原位点 或未经确认，将进行此类任务。 (5) 不同胶原蛋白基因研究之间的遗传距离 (1)和(2)以及选定的同线基因座将被确定。 这些研究的目的有四个：(i) 识别和 表征胶原蛋白基因的突变，这些突变负责 某些结缔组织家族性疾病，(ii) 确定 这些突变对胶原蛋白 mRNA 和蛋白质合成的影响 结构； (iii) 识别结构的正常变化以及 限制性片段揭示的胶原蛋白基因的组织 长度多态性； (iv) 增加我们对人类基因图谱的了解， 就染色体定位和遗传距离而言 紧密相连的基因座和各种胶原蛋白基因之间的关系。

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.