Synthesis of 125I-labeled N-3-(4-hydroxyphenyl)propionyl aminoacyl transfer ribonucleic acids.
A method for the isolation and labeling to high specific radioactivity of individual isoaccepting tRNAs is described. After blocking reactive minor bases by acetylation and iodination of the crude tRNA, a single family of isoacceptors was aminoacylated. Individual isoacceptors were separated by chromatography on RPC-5 and then acylated with the 3-(4-hydroxyphenyl)propionyl ester of N-hydroxysuccinimide. The product was purified by chromatography on BD-cellulose and RPC-5. This derivatized tRNA was then iodinated with 125I- and Chloramine-T to give a product containing between 5 X 10(7) and 3 X 10(8) dpm/microgram. The suitability of such labeled tRNAs for hybridization to homologous DNA in solution and cytological preparations of chromosomes is discussed with particular reference to Drosophila melanogaster.
D002850:Chromatography, Gel; D007457:Iodine Radioisotopes; D007553:Isotope Labeling / Q000379:methods; D012346:RNA, Transfer, Amino Acyl / Q000302:isolation & purification
D007457:Iodine Radioisotopes; D012346:RNA, Transfer, Amino Acyl