COLLAGEN GENES IN HERITABLE CONNECTIVE TISSUE DISORDERS

我的总体目标是获得教学培训和实践 实验室经验成为独立的生物医学 调查员 我打算把这种训练应用到研究 胶原蛋白基因的改变对各种遗传性疾病的影响， 包括选定的纤维结缔组织遗传性疾病， 软骨营养不良 建议采取以下办法： (1)这些细胞中各种胶原蛋白基因的结构和组织 通过Southern印迹分析确定疾病。 不寻常的模式 将与对照组中的结果进行比较，以确定 前者是基因突变的结果。 (2)胶原蛋白基因结构的更微妙的改变将是 通过连锁分析发现。 多个受影响的大家庭 将检查个体的“突变”表型的共分离 和一个特定的胶原蛋白等位基因，其遗传通过多态性检测， 限制性位点。 (3)在（1）或（2）表明突变的个体或家族中 在胶原基因中，对该基因编码的mRNA和蛋白质的研究将 进行研究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)对于那些染色体分配未知的胶原基因座 或未经确认，将进行此类分配。 (5)研究中各种胶原基因之间的遗传距离， (1)（2）选择同线基因座。 这些研究的目的有四个方面：（一）查明和 表征胶原蛋白基因中的突变， 某些家族性结缔组织疾病，（ii）确定 这些突变对胶原mRNA和蛋白质合成的影响， （iii）确定结构中的正常变化， 限制性片段显示胶原基因的组织 长度多态性;（iv）增加我们对人类基因图谱的了解， 从染色体定位和遗传距离两个方面来看 紧密连锁的基因座和各种胶原基因之间的联系。

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.