GENETIC STUDIES OF NON-ERYTHROID SPECTRIN

我们的长期目标是定义特征细胞和 幽灵蛋白的发育作用或作用，并了解如何 幽灵蛋白分子对特定的细胞功能有贡献。我们 我相信我们的长期目标可以通过使用模型来实现 系统，果蝇黑腹果蝇，可以在这两个 利用细胞和组织水平的组合 生物化学、结构和遗传方法。 将使用抗体来研究其分布和表达 在细胞培养和发育过程中的幽灵蛋白 鉴定表达果蝇血影蛋白的DNA序列。克隆人 这些DNA序列将被用来绘制物理位置 果蝇遗传图谱上的血影蛋白编码序列 经典的遗传分析和体外突变生产将 然后被执行。这些突变体将被用于研究 血影蛋白在体内的功能，并确定其影响 完全缺乏功能性的血影蛋白。克隆人也将是 与反义抑制技术结合使用 表达抑制培养细胞中的幽灵蛋白表达，从而 我们可以探索功能幽灵蛋白缺失的影响。 突变的幽灵蛋白在培养细胞中表达的后果。 幽灵蛋白存在于几乎所有人类细胞中，但健康细胞除外。 有缺陷的幽灵蛋白表达的后果只是 在红血球中发现。建议的实验应该是 深入了解血影蛋白在非 红系细胞。

Our long term goal is to define the characteristic cellular and developmental role or roles of spectrin and understand how spectrin molecules contribute to specific cellular functions. We believe our long term goal can be attained by using a model system, Drosophila melanogaster, that can be investigated at both the cellular and the tissue level using a combination of biochemical, structural and genetic approaches. Antibodies will be used to study the distribution and expression of spectrin both in cell culture and during development and to identify DNA sequences that express Drosophila spectrin. Clones of these DNA sequences will be used to map the physical location of the spectrin coding sequences on the genetic map of Drosophila and classical genetic analysis and in vitro mutant production will then be carried out. The mutants will be used to investigate spectrin function in vivo and to determine the effects of the complete absence of functional spectrin. The clones will also be used together with the technology of anti-sense inhibition of expression to inhibit spectrin expression in cultured cells so that we may probe the effects of the absence of functional spectrin or the consequences of mutant spectrin expression in cultured cells. Spectrin is present in nearly all human cells but the health consequences of defective spectrin expression have only been explored in erythrocytes. The proposed experiments should provide an in-depth understanding of spectrin's role in non- erythroid cells.