The proposed research program will focus on the development of efficient, chemical ways to introduce biotin into synthetic DNA fragments, that can subsequently be detected with the streptavidin-alkaline phosphatase signaling system. This nonradioactive DNA hybridization probe has several advantages over the radioactive labeling techniques, including much higher shelf life, greater convenience and safety, high detectability and automation capability. Such synthetic biotinylated oligonucleotides of mixed sequences, can be used directly as hybridization probes for the identification and isolation of specific genes, in a very short period of time. The sensitivity of the method for visual detection of the hybridized probes, is sufficiently high that 0.5 fem to mole lO-l5 mole) of plasmid DNA can be detected. The development of these biochemical probes will have significant impact on biomedical research and may find applications in the diagnosis of genetically determined conditions, such as viral diseases and cancers. The chemical synthesis of biotinylated DNA fragments can be carried out by the solid phase phosphoramidite method. The feasibility of this approach will be examined in the proposed research. While the original focus of this work will be on oligonucleotides labeled with biotin, it is possible to subsequently examine several other kinds of nonradioactive labeling systems (fluorescent probes, electron spin resonance probes, photoaffinity labels, etc.), with potential applications in biochemical research.Anticipated Results and
Potential Commercial Applications: The potential commercial application as described by the awards Techniques developed in the research could be applied to cancer diagnosis, clinical diagnosis, and diagnosis of viral diseases.
