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Sequence analysis of transfer RNA 7 ser of Drosophila melanogaster Cribbs, David Lamar

Abstract

Sequence analysis of tRNA 7 ser from Drosophila melanogaster was carried out, primarily by the formamide degradation and post-labeling method of Stanley and Vassilenko. Preliminary analysis was on the terminal nucleoside-5', 3'-bis [51 - ³²P] phosphates of electrophoretically separated [51 - ³²P] ribooligonucleotides, identifying the [³² P]-nucleotides by chromatography on PEI-cel1ulose plates. Further analysis of possible modified nucleotides was performed by thin layer chromatography of [5' -³²P] nucleoside phosphates derived by nuclease P-j digestion from [51 - ³²P] oligomers. The partial sequence generated in this fashion was supplemented by ladder gel sequence analysis of [51 - P]tRNA ser , and to a limited extent by two dimensional homochromatography. In this way, a sequence was obtained that is complete except for part of the aminoacyl stem and the 5'-end of the extra arm. The data are consistent with a sequence for tRNAser of pGCAGmUUGUGGCac4CGAGCGmGDDAAGGCXUCUGA— m3CUI GAi 6AAujCAGAUmUCCCUm3CUGGGAGm5CGUAGGTijjCGm1 AAUCCUACCGACUGCNCCA.

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