Stable isotope enrichment in synthetically prepared oligonucleotides coupled with multi-dimensional NMR techniques will greatly enhance the sensitivity and resolution of this technique when applied to the study of RNA. This enhancement of the NMR technique will enable a rapid and easy determination of protein RNA complexes and protein RNA interaction in solution. These studies are important for the design of new pharmaceuticals and drug targeting. Uniformly labeled (with 2H, 13C, and 15N) ribonucleosides are presently unavailable. The goal of this research is to study the production and determine the economic feasibility of producing 2H-, 13C-, and 15N- labeled ribonucleosides. It is proposed to produce four common ribonucleosides using yeast, a Candida species, which are widely reported to produce up to 15 percent of its biomass as RNA. The hydrolyzed, stable isotopically labeled algal biomass will be used as the production feed stock. Success of this research will establish the basis for the production of stable isotopically labeled oligonucleotide that are vital for the study of protein-RNA interactions.
