Our goals are to develop novel physical methodology for rapid mapping of the human genome and further studying its organization. Toward this end, the physical basis of pulsed field gradient gel electrophoresis (PFG) will be investigated with an aim toward greatly increasing its present size resolution. New spectroscopic-electrophoretic methods will be developed for studying the molecular dynamics of electrophoretic behavior of large DNA molecules in a gel matrix as well as to provide novel means of DNA analysis by size or sequence. Additionally, new physical methods potentially capable of fractionating nucleoprotein complexes and also producing human karyotypes will be developed. This new methodology, termed induced anisotropic filtration (IAF), could be intrinsically capable of preparative chromosome fractionation, which would facilitate and speed human gene mapping. In fact, if IAF is successful, it could become a major human diagnostic tool. Our long-term goal is to develop a complete methodology capable of lifting chromosomal regions defined by chromosomal polymorphisms and finding the particular mutation rapidly and easily. Rapid characterization of deletions and rearrangements in relatively large chromosomal regions not visible cytologically and too difficult to find by chromosome walking will be pursued. In short, a physical biological methodology will be developed making human genome analysis vastly more accessible to human geneticists and making the molecular basis of diseases more discernable.
