MOLECULAR ANALYSIS OF CELL DEATH GENES

虽然所有细胞最终都会死亡，但人们对这种分子知之甚少。 导致这种发育现象的机制。一个非常棒的 吸引人的模型系统用于研究，是节间肌肉(ISM)的 六纹夜蛾成虫后36小时内死亡 浮现。ISM对堕落的承诺涉及 激活了一组新的细胞死亡基因，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 洞察这些基因在昆虫和昆虫中的身份和功能 脊椎动物。将使用一系列分子方法来分离， 对细胞死亡基因进行测序和表达。此外，培养细胞和 昆虫肌肉纤维将被注射或转基因其中一种 以下是我们的克隆产生的分子：反义转录本， 正义转录本、抗体、纯化的表达蛋白或重组体 由融合到假定的细胞死亡基因启动子的报告基因制成。 这些实验应该会极大地增加我们对 细胞用来在发育和衰老过程中死亡的分子。 我目前是动物学助理教授，背景是 生理学、膜生物物理学和最近的分子生物学。我的 教学和行政承诺相当苛刻，这限制了 我既能学习新技术，又能参观其他实验室进行培训。 RCDA提供的时间将极大地提高我的能力 来开发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 调节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.