CHARACTERIZATION AND EXPRESSION OF TUBULIN GENES

有鞭毛的单细胞真核生物中的微管蛋白基因家族， 莱茵衣藻，由两个β微管蛋白基因组成，编码 相同的蛋白质，和两个α微管蛋白基因编码轻微 不同的产品。 为什么这种单倍体生物保持重复的基因 以及基因在生命中扮演着什么样的角色 循环，是需要解决的问题。 抗性突变体的分离 抗微管除草剂已经确定了微管蛋白和非微管蛋白， 微管蛋白基因可能对微管组织很重要， 功能 分子生物学和遗传学技术将用于 了解这些基因的功能。 抗- 将表征微管药物。 分布的比较， 稳定性和组装特性的突变体和野生型 型细胞，在存在和不存在抗微管药物的情况下， 将通过使用免疫荧光技术和通过检查 鞭毛的组装和拆卸。 条件致死突变体 检查以确定它们被阻断的细胞周期阶段。 将检查基因在染色体传递中的作用。 基因技术将被用来识别其他基因， 与细胞中的α微管蛋白相互作用。 分离鉴定的基因 通过突变分析将使用转化来完成， 用克隆的衣原体基因补充衣原体突变。 转座子诱变也将被开发作为基因工程的工具。 隔离 转化将用于鉴定顺式作用序列 转录化学治疗剂的重要元件。 的 拟议的研究将提供深入了解抗微管蛋白的作用， 药物，以及更普遍的重要问题， 在真核系统中参与微管功能的基因。

The tubulin gene family in the flagellated unicellular eukaryote, Chlamydomonas reinhardtii, consists of two beta tubulin genes coding for identical proteins, and two alpha tubulin genes coding for slightly different products. Why this haploid organism maintains duplicate genes for the tubulins, and what rules are played by the genes in the life cycle, are questions to be addressed. The isolation of mutants resistant to anti-microtubule herbicides has identified both tubulin and non- tubulin genes that may be important for microtubule organization and function. Techniques of molecular biology and genetics will be used to understand the functions of these genes. Mutant lines resistant to anti- microtubule drugs will be characterized. Comparison of the distribution, stability, and assembly properties of microtubules in mutant and wild- type cells, in the presence and absence of the anti-microtubule drugs, will be obtained by using immunofluorescence techniques and by examining flagellar assembly and disassembly. Conditional-lethal mutants will be examined to determine the cell cycle stage at which they are blocked. The role of the genes in chromosome transmission will be examined. Genetic techniques will be used to identify other genes whose products interact with alpha tubulin in the cell. Isolation of genes identified by mutational analysis will be accomplished using transformation to complement Chlamydomonas mutations with cloned Chlamydomonas genes. Transposon mutagenesis will also be developed as a tool for gene isolation. Transformation will be used to identify cis-acting sequence elements important for transcriptional chemotherapeutic agents. The proposed research will provide insight into the action of anti-tubulin drugs, as well as the more generally important question of identifying genes involved in microtubule function in eukaryotic systems.