NEW TECHNIQUES FOR COMPLEX MACROMOLECULAR ASSEMBLIES

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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.