CUTICULAR COLLAGEN ASSEMBLY IN C ELEGANS

线虫的角质层胶原蛋白组装

• 批准号: 2732802
• 负责人: JENNIFER R CREW
• 金额: $ 2.92万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2732802
• 关键词: Caenorhabditis elegans; collagen; gene expression; genetically modified animals; invertebrate cuticle; northern blottings; phenotype; protein structure function; western blottings

The cuticle of the nematode Caenorhabditis elegans is a multi-layered collagenous "skeleton' that plays crucial roles in both morphogenesis and motility. Because collagen is a major constituent of the cuticle, genetic analysis of cuticular morphology is useful in studying the structure and function of collagen molecules in vivo. The experiments in this proposal concentrate on analyzing the structure and assembly of one substructure within the cuticle, using the bli-1 and bli-2 genes. The cuticles of animals mutant in these genes are covered by fluid-filled blisters, which result from an apparent separation between cuticular layers. Animals mutant for bli-1 have been shown to lack structures referred to as "struts", which function to hold two cuticular sublayers together. The function of these struts may be analogous to the function of anchoring fibers in vertebrate skin; these collagenous structures bind together the dermis and epidermis. The experiments outlined in this proposal aim first to use a variety of genetic and molecular biological techniques to characterize the genomic sequences corresponding to these loci. The next steps in this outline attempt to analyze the loci in terms of RNA and protein expression patterns using molecular and immunological techniques as well as gene fusion studies in transgenic animals. These experiments will also further investigate the phenotype of animals mutant in bli-1 and bli-2 using genetic and ultrastructural analysis. The final set of experiments in this study are aimed at gaining an understanding of the morphogenesis of the struts using a combination of classical genetic techniques as well as transgenic animal technology.

秀丽隐杆线虫的角质层是一个多层的 胶原“骨架”，在形态发生和 能动性由于胶原蛋白是角质层的主要成分，遗传 表皮形态学的分析有助于研究结构， 胶原蛋白分子在体内的功能。该提案中的实验 集中分析一个子结构的结构和装配 在角质层内，使用bli-1和bli-2基因。的cuberance 这些基因突变的动物被充满液体的水泡覆盖， 这是由表皮层之间明显的分离造成的。动物 BLI-1突变体已显示缺乏称为 “支柱”，其功能是将两个表皮亚层保持在一起。的 这些支柱的功能可能类似于锚定的功能 脊椎动物皮肤中的纤维;这些胶原结构将 真皮和表皮。 本提案中概述的实验旨在首先使用各种 基因和分子生物学技术来表征基因组 对应于这些基因座的序列。本大纲中的后续步骤 试图从RNA和蛋白质表达的角度分析基因座 使用分子和免疫学技术以及基因 在转基因动物中的融合研究。这些实验还将进一步 研究动物bli-1和bli-2突变的表型， 遗传和超微结构分析。最后一组实验， 本研究的目的是获得一个了解的形态发生， 同时结合经典的遗传技术 转基因动物技术。