| Research Abstract |
This research project has focussed on a new technology for gene diagnosis. Several fatal diseases such as AIDS, ATL, cancer, and so on are regarded as genetic diseases and it is hard to detect them before their symptoms appear. Among methods for gene diagnosis, DNA-probe methodology was thought to be very suitable for such diseases and adopted in this research. The methodlogy is based on the nucleic acid chemistry and gene informations so far reported. As a tool for DNA-probe methodology, several nucleic acid analogs labeled with functional groups were synthesized. They are fluorescent-labeled nucleic acid analogs(F-DNA-probe)and electrospin-labeled ones(S-DNAprobe). When the former analogs were used, measured were fluorescence depolarization spectra, which sensitively reflect the molecular motion offluorescent materials, namely of nucleic acid analogs. For the latter, ESR spectra change, especially line-width change, were monitored. When F-DNA-probe was mixed with its complementary oligonucleotide, anisotropy of fluorescence changed remarkably and the change was molecular weight dependent until 10000. When S-DNA-probe was mixed with its complementary oligonucoleotide, the line-width of the ESR spectrum appreciably increased. These results indicate that both DNA-probes are suitable to detect the hybrid formation in solution phase, namely the presence of target genes.
In order to demonstrate the ability of such probes to detect natural DNAs, M13-phage DNA as well as the deletion mutant of M13-ohage DNA were chosen. The probes efficiently distinguished the two DNAs depending on their sequences. These results suggest that suitably labeled nucleic acids can be used as tools to detect target DNA in solution using fluorescence depolarization spectrum and electron spin resonance spe : ctrurb. And it is promising that these methodology can be used for automatic machine for gene diagnosis.
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