CHARACTERIZATION AND EXPRESSION OF TUBULIN GENES

微管蛋白基因的表征和表达

• 批准号: 3279094
• 负责人: CAROLYN D SILFLOW
• 金额: $ 13.99万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3279094
• 关键词: Chlorophyta; drug resistance; eukaryote; flagellum; gene expression; gene mutation; genetic manipulation; herbicides; immunofluorescence technique; laboratory rabbit; lethal genes; microtubules; molecular cloning; mutant; nucleic acid sequence; plant genetics; protein biosynthesis; regulatory gene; transposon /insertion element; tubulin

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.

鞭毛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阿尔法微管蛋白基因编码 不同的产品。为什么这种单倍体生物体保持重复的基因 对于微管蛋白，以及生命中的基因所起的作用 循环，都是需要解决的问题。抗病突变体的分离 抗微管除草剂已确定微管蛋白和非微管除草剂 微管蛋白基因可能对微管组织和 功能。将使用分子生物学和遗传学技术来 了解这些基因的功能。抗赤霉病突变系 将对微管药物进行表征。比较分布情况， 突变体和野生型微管的稳定性和组装特性 类型细胞，在有无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。条件致命性突变体将是 检查以确定它们被阻断的细胞周期阶段。 我们将研究这些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的产物 与细胞内的α-微管蛋白相互作用。已鉴定基因的分离 通过突变分析将使用转换到 用克隆的衣藻基因补充衣藻突变。 转座子突变也将作为基因检测的工具而被开发。 与世隔绝。将使用转换来识别顺式作用序列 转录型化疗药物的重要成分。这个 拟议的研究将为了解抗微管蛋白的作用提供洞察 毒品，以及更普遍的重要的识别问题 涉及真核系统中微管功能的基因。