NEW TECHNIQUES FOR COMPLEX MACROMOLECULAR ASSEMBLIES

该计划的总体目标是开发新技术， 研究大分子和组装。 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， 用于在体内产生标记蛋白的分子和方法。 脉冲场 梯度凝胶电泳允许DNA的高分辨率分离 分子大小为5000 kb。 这将用于开发快速基因 分离和作图方法。 电流脉冲场的改进 技术和方法的特定片段较大的DNA将是 开发，以便快速映射方法可以扩展到人类 基因. 其中提出的应用是电泳核型， 基因组指纹，探索邻近基因组结构的技术， 一种克隆的标记，以及探索移动的遗传动力学的方法， 元素 在其他研究中，链霉亲和素和水母发光蛋白的基因将被 克隆并在哺乳动物细胞中表达。 将构建向量， 产生这些蛋白质和正常细胞蛋白质之间的融合。 这 将允许任何基因产物的特异性体内标记。

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.