THE GENETICS OF MICROTUBULE ASSEMBLY AND FUNCTION

微管是一种重要的细胞器。 并且功能多样。此外，不同的MT是不同的 对不同解聚剂敏感，结合不同MT 相关蛋白质。据推测，MT多样性可能是 这是由于MTS的微管蛋白组成的差异造成的。微管蛋白 MTS的主要结构蛋白由α和α两个亚基组成 β微管蛋白。这些亚基中的每个都有多个基因 已经被检查过的生物体。不仅有基因 同种类型的α和β微管蛋白，但也有 转录后修饰的微管蛋白亚型，其中一些已经被 被证明局限于特定的MT人群。在此应用程序中，我们 建议分析微管蛋白基因亚型与MT的关系 丝状真菌尼杜拉曲霉的结构和功能。 四个曲霉微管蛋白基因中的三个，两个β微管蛋白和一个 α微管蛋白已被克隆和测序。贝塔微管蛋白已经 通过DNA介导的整合转化定位到它们的遗传位点， 其中一个基因的功能已经通过dna载体进行了检测。 “基因破坏。”我们建议克隆剩余的α-微管蛋白基因， 利用DNA介导的整合转化对两者进行鉴定 未分配的α-微管蛋白序列的遗传位点和使用“基因” 和体外诱变，以检测所有的功能 这些基因。此外，我们建议使用同型特异性抗微管蛋白 利用荧光技术定位特异性微管蛋白的抗体 免疫细胞化学和构建新型微管蛋白基因以确定 微管蛋白基因同型是否可以互换。

Microtubules (MTs) are important cellular organelles that are structurally and functionally diverse. Moreover, different MTs are differentially sensitive to different depolymerizing agents and bind different MT associated proteins. It has been hypothesized that MT diversity may be caused by differences in the tubulin composition of MTs. Tubulin, the major structural protein of MTs, is composed of two subunits, alpha and beta tubulin. There are multiple genes for each of these subunits in all of the organisms that have been examined. Not only are there genetic isotypes of alpha and beta tubulin, but there are also posttranscriptionally modified tubulin isotypes, some of which have been shown to be localized to specific MT populations. In this application we propose to analyze the relationship between tubulin genetic isotypes and MT structure and function in the filamentous fungus Aspergillus nidulans. Three of the four Aspergillus tubulin genes, two beta tubulins and one alpha tubulin have been cloned and sequenced. The beta tubulins have been assigned to their genetic loci by DNA mediated integrative transformation, and the function of one of these genes has been examined by DNA mediated "gene disruption." We propose to clone the remaining alpha tubulin gene, to use DNA mediated integrative transformation to assign the two as yet unassigned alpha tubulin sequences to their genetic loci and to use "gene disruption" and in vitro mutagenesis to examine the functions of all of these genes. In addition we propose to use isotype specific antitubulin antibodies to localize specific tubulin to specific MTs by fluorescence immunocytochemistry and to construct chaemeric tubulin genes to determine whether tubulin genetic isotypes are interchangeable.