STRUCTURAL STUDIES OF DROSPHILA MIRANDA PROTEIN

不对称细胞分裂是一种在发育中的生物体中实现细胞多样性的现象。了解细胞不对称分裂的分子机制对于理解异常过程（如肿瘤发生和发育缺陷的起源）具有根本的重要性。在果蝇成神经细胞中，这些机制的核心是米兰达，一种在有丝分裂期间负责细胞命运决定因素不对称分离的支架蛋白。米兰达及其与货物蛋白Prospero和Staufen的复合物的结构将利用X射线晶体学技术和方法进行研究。将使用采用构建体筛选策略的合理方法获得适量的可溶性、纯的、稳定的、单多样性材料。随后的结晶试验将基于稀疏矩阵阶乘策略。使用这种“双屏”方法有助于以有利的方式获得衍射质量的晶体，从而可以实现结构测定。

Asymmetric cell division is a phenomenon by which cellular diversity is achieved in the developing organism. Knowledge of the molecular mechanisms by which asymmetric cell division normally occurs is of fundamental importance and relevant to understanding aberrant processes such as tumorigenesis and the origin of developmental defects. Central to these mechanisms in the Drosophila neuroblast is Miranda, a scaffold protein responsible for the asymmetric segregation of cell-fate determinants during mitosis. The structure of Miranda and its complexes with cargo proteins Prospero and Staufen will be investigated utilizing X- ray crystallographic techniques and methodologies. Appropriate quantities of soluble, pure, stable, monodiverse material will be obtained using a rational approach that employs a construct screening strategy. Subsequent crystallization trials will be based on sparse matrix factorial strategies. Use of this "double-screen" approach serves to obtain diffraction quality crystals in an expedient manner so that structure determination may be achieved.