NEW TECHNIQUES FOR COMPLEX MACROMOLECULAR ASSEMBLIES

该计划的总体目标是开发新技术，以 研究大分子和组装体。目前的具体目标是 开发处理和分析染色体大小的DNA的方法 用于在体内生产标记蛋白质的分子和方法。脉冲场 梯度凝胶电泳法可实现DNA的高分辨率分离 大到5000 kb的分子。这将被用来开发快速基因 隔离和作图方法。电流脉冲场的改进 较大DNA的特定片段的技术和方法将是 开发出了快速映射方法，可以扩展到人类 基因。在建议的应用中，包括电泳核型和 基因组指纹，探索邻近基因组结构的技术 一种克隆标记和探索移动遗传动态的方法 元素。在其他研究中，链霉亲和蛋白的基因将是 克隆并在哺乳动物细胞中表达。向量将被构造为 在这些蛋白质和正常细胞蛋白质之间产生融合。这 将允许对任何基因产品进行特定的体内标记。

The general goal of this program is the development of new techniques to study macromolecules and assemblies. The current specific objectives are to develop methods for handling and analysis of chromosome-sized DNA molecules and methods for producing labeled proteins in vivo. Pulsed field gradient gel electrophoresis allows high resolution separations of DNA molecules as large as 5000 kb. This will be used to develop rapid gene isolataion and mapping methods. Refinements of current pulsed field techniques and methods for specific fragmentation of larger DNA will be developed so that the rapid mapping methods can be extended to human genes. Among the proposed applications are electrophoretic karyotypes and genome finger prints, techniques for exploring genome structure adjacent to a cloned marker, and methods for exploring the dynamics of mobile genetic elements. In other studies the genes for strepavidin and aequorin will be cloned and expressed in mammalian cells. Vectors will be constructed that produce fusions between these proteins and normal cellular proteins. This will allow specific in vivo labeling of any gene product.