CUTICULAR COLLAGEN ASSEMBLY IN C ELEGANS

线虫的角质层胶原蛋白组装

• 批准号: 2442775
• 负责人: JENNIFER R CREW
• 金额: $ 2.44万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2442775
• 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.

线虫线虫的角质层是多层的 在形态发生和生长发育中起关键作用的胶原性“骨骼” 能动性。因为胶原蛋白是角质层的主要成分，遗传 角质层形态分析有助于研究其结构和功能。 胶原分子在体内的功能。这项提案中的实验 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。皮肤的角质层 这些基因突变的动物被充满液体的水泡覆盖，这 这是由于角质层之间的明显分离造成的。动物 BLI-1的突变体被证明缺乏被称为 “支撑物”，其功能是将两个角质层亚层保持在一起。这个 这些支柱的功能可以类似于锚固的功能 脊椎动物皮肤中的纤维；这些胶原结构将 真皮和表皮。 这项提案中概述的实验首先旨在使用各种 基因组特征的遗传和分子生物学技术 与这些基因座相对应的序列。本大纲中的下一步 试图从RNA和蛋白质表达的角度分析这些基因座 使用分子和免疫学技术以及基因的模式 转基因动物的融合研究。这些实验还将进一步 BLI-1和BLI-2基因突变动物的表型研究 遗传和超微结构分析。的最后一组实验 这项研究的目的是为了了解其形态发生机制。 Struts也使用了经典基因技术的组合 作为转基因动物技术。