RUI: A Genetic Analysis of Conjugation in Tetrahymena thermophila

Cole 9807555淡水纤毛虫四膜虫嗜热四膜虫将作为实验生物体，用于探索接合程序的发育规律。研究的目标是：1)进一步探索嗜热四膜虫接合的发育依赖关系和发育逻辑；2)利用先进的显微技术，对发育关键点期间的核-细胞骨架相互作用进行超详细的描述；3)协助开发和/或实施分子遗传学程序，旨在识别接合特异性基因并探索其功能。嗜热四膜虫的接合是有趣的，因为减数分裂、核交换和受精、程序性核消除、合子基因组的转录激活和基因组重组等基本细胞事件都发生在一个高度同步的种群中，在12小时的时间表内。在前一个奖项的任期内，开发了一种产生影响接合途径的突变的方法。分离并鉴定了13个几乎影响到配对发育各个阶段的接合突变体。这样的特征使得可以产生一个详细的发育依赖模型，这是共轭计划的基础。这项工作的继续不仅将允许进一步深入了解基本的遗传机制，而且将有助于教育学生基础研究的需求和满足感。

Cole 9807555The freshwater ciliate, Tetrahymena thermophila, will be the experimental organism used to explore the developmental regulation of the conjugation program. The research objectives are: 1) To further explore the developmental dependencies and developmental logic underlying conjugation in Tetrahymena thermophila, 2) To use advanced microscopy techniques to produce an ultra-detailed description of nuclear-cytoskeletal interactions during critical points in development and, 3) To assist in the development and/or implementation of molecular genetic procedures designed to identify conjugation specific genes and explore their function. Conjugation in Tetrahymena thermophila is interesting in that such fundamental cellular events as meiosis, nuclear exchange and fertilization, programmed nuclear elimination, transcriptional activation of the zygotic genome and genomic reorganization all occur within a highly synchronous population at well defined times during a twelve hour schedule. A method of generating mutations that affect the conjugation pathway was developed during the tenure of the previous award. Thirteen conjugation mutants affecting nearly every stage of conjugal development were isolated and characterized. Such characterization allowed the production of a detailed model of developmental dependencies, which underlie the conjugation program. The continuation of this work will not only allow further insight into fundamental genetic mechanisms but will serve to educate students in the demands as well as the satisfaction of basic research.