ORGANIZATION OF THE MAMMALIAN MITOTIC SPINDLE

描述：本申请中提出的实验目标 是鉴定和功能表征结构蛋白， 通过以下具体目标构成主轴矩阵： L。使用高度丰富的无细胞有丝分裂酯组装系统 在有丝分裂纺锤体基质的成分中，研究人员将 制备特异性识别三种主要蛋白的抗体 有丝分裂的纺锤体基质。 2.使用针对特定目标L制备的抗体，调查员将 使用免疫耗竭和微量注射来确定这些 蛋白质是有丝分裂纺锤体组装所必需的。 3.利用针对特定目标制备的抗体，L将分离cDNA. 编码有丝分裂纺锤体基质的每种蛋白质的克隆 确定它们的主要氨基酸序列。

DESCRIPTION: The objective of the experiments proposed in this application are to identify and functionally characterize the structural proteins that comprise the spindle matrix through the following specific aims: l. Using a cell-free mitotic aster assembly system that is highly enriched in the components of the mitotic spindle matrix, the investigator will prepare antibodies that specifically recognize the three major proteins of the mitotic spindle matrix. 2. Using the antibodies prepared in Specific Aim l, the investigator will use immunodepletion and microinjection to determine if each of these proteins is required for the assembly of the mitotic spindle. 3. Using the antibodies prepared in Specific Aim l, he will isolate cDNA clones encoding each of these proteins of the mitotic spindle matrix and determine their primary amino acid sequence.