MOLECULAR ANALYSIS OF CELL DEATH GENES

虽然所有的细胞最终都会死亡，但人们对细胞的分子结构知之甚少。 导致这种发展现象的机制。 一个非常 有吸引力的模型系统的研究，是节间肌肉（ISM）的 天蛾Manduca sexta，在成虫后36小时内死亡， 出现。 ISM对退化的承诺涉及到 一组新的细胞死亡基因的激活，其中五个已经被 在我的实验室里克隆的 本提案中概述的实验将提供 深入了解这些基因在昆虫和 脊椎动物 一系列的分子方法将被用于分离， 测序并表达细胞死亡基因。 此外，培养细胞和 昆虫肌肉纤维将被注射或转染一种 从我们的克隆中产生了以下分子：反义转录物， 有义转录物、抗体、纯化的表达蛋白或重组体 用与假定的细胞死亡基因启动子融合的报告基因制造。 这些实验应该大大增加我们对 细胞在发育和衰老过程中死亡所用的分子。 我目前是动物学的助理教授，有着 生理学、膜生物物理学和最近的分子生物学。 我 教学和行政工作要求很高，这限制了 我学习新技术和参观其他实验室进行培训的能力。 RCDA提供的时间将大大提高我的能力 来利用这个强大的细胞死亡模型系统。 很明显 对这些模型系统的检查将提供对 介导动物细胞衰老和死亡的分子机制。

While all cells ultimately die, very little is known about the molecular mechanisms which bring about this developmental phenomenon. One very attractive model system for study, is the intersegmental muscles (ISM) of the moth Manduca sexta, which die during the 36 hours following adult emergence. The commitment of the ISM to degenerate involves the activation of a new set of cell death genes, five of which have been cloned in my lab. The experiments outlined in this proposal will provide insight into the identity and function of these genes in both insects and vertebrates. A range of molecular approaches will be used to isolate, sequenced and express cell death genes. Additionally, culture cells and insect muscle fibers will be injected or transfected with one of the following molecules generated from our clones: anti-sense transcripts, sense transcripts, antibodies, purified expressed proteins or recombinants made with a reporter gene fused to putative cell-death gene promotors. These experiments should greatly increase our understanding of the molecules which are used by cells to die during development and aging. I am currently an assistant professor of Zoology, with a background in physiology, membrane biophysics and recently, molecular biology. My teaching and administrative commitments are quite demanding, which limits my ability to both learn new techniques and visit other labs for training. The time made available by an RCDA will dramatically increase my ability to exploit this powerful cell death model system. It is clear that examination of such model systems will provide basic insight into the molecular mechanisms which mediate senescence and death in animal cells.