Tubulin Gene Expression During Morphogenesis of Arabidopsis

拟议研究的长期目标是了解 高等植物生长发育的遗传控制。的 研究的重点是形态发生的遗传控制， 模式植物拟南芥。的主要目标 本研究旨在确定个体的具体角色， 微管复合体中的微管蛋白基因和微管蛋白亚型 主要参与细胞分裂和细胞分裂的过程 控制植物组织形态发生的伸长， 机关拟议调查的具体目的是：1） 来描述表达的时空模式 通过北方印迹和原位杂交技术 杂交实验，2）定位，空间和 在时间上，发育中的植物中的个别微管蛋白亚型 组织和器官的蛋白质印迹分析， 用同种型特异性的免疫荧光显微镜进行 抗体，3）使用免疫荧光显微镜和同种型- 特异性抗体，以确定哪些亚型存在于 含有微管的结构如前期带， 有丝分裂纺锤体、成膜体和皮层阵列，4）至 鉴定顺式作用序列和反式作用蛋白， 控制拟南芥组织特异性表达的基因 微管蛋白基因和5）利用“过表达的”反义基因 由农杆菌产生的转基因植物中的特异性序列 根癌农杆菌Ti质粒介导的转化，以探测 个别微管蛋白基因的功能。 %%% 微管是一种丝状聚合物， 高等生物细胞的几个基本过程。在 植物细胞，它们参与细胞分裂和细胞 扩张，植物形态发生的关键过程。这个项目是 代表了一个详细的分子研究的组成部分， 植物中的微管

The long-term goal of the proposed research is to understand the genetic control of growth and development of higher plants. The investigation focuses on the genetic control of morphogenesis in the model plant Arabidopsis thaliana. The major objective of the present study is to determine the specific roles of individual tubulin genes and tubulin isoforms in the microtubule complexes centrally involved in the processes of cell division and cell elongation that govern the morphogenesis of plant tissues and organs. The specific aims of the proposed investigations are 1) to characterize the spatial and temporal patterns of expression of individual tubulin genes by northern blot and in situ hybridization experiments, 2) to localize, spatially and temporally, the individual tubulin isoforms in developing plant tissues and organs by western blot analyses and immunofluorescence microscopy carried out with isoform-specific antibodies, 3) to use immunofluorescence microscopy and isoform- specific antibodies to determine which isoforms are present in microtubule containing structures such as preprophase bands, mitotic spindles, phragmoplasts, and the cortical arrays, 4) to identify the cis-acting sequences and trans-acting proteins and genes that control the tissue specific expression of Arabidopsis tubulin genes and 5) to utilize "overexpressed" antisense gene specific sequences in transgenic plants produced by Agrobacterium tumefaciens Ti plasmid mediated transformation to probe the function of individual tubulin genes. %%% Microtubules are filamentous polymers that play central roles in several basic processes of the cells of higher organisms. In plant cells they are involved in cell division and cell expansion, processes key to plant morphogenesis. This project is represents a detailed molecular study of the components of microtubules in plants.