Separation and characterization of some ribonucleic acids from imaginal discs of Drosophila melanogaster
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The RNA obtained from mass isolated imaginal discs of Drosophila melanogaster cultured in vitro in 3H-5-uridine has been studied. A separation procedure employing benzoylated, naphthoylated DEAE cellulose column chromatography was used to obtain several fractions of RNA, including one consisting primarily of ribosomal RNA, and three others relatively free of rRNA and transfer RNA. Other than its behavior in BND-cellulose chromatography, the RNA fractions have been examined with the following procedures: (1) sucrose gradient centrifugation, (2) heat labilities with apparent shifts in S-values, (3) changes in apparent secondary structure due to monovalent cation concentration, (4) Cs2SO4−CsCl equilibrium gradient centrifugation, (5) dimethyl sulfoxide gradient centrifugation, (6) “melting out” studies in different salt (Na+) concentrations, (7) limited digestion with different kinds of RNase, (8) kinetics of labeling, and (9) DNA-RNA hybridization. Marked differences were found among the RNA fractions using these techniques. The addition of β-ecdysone to the cultured imaginal discs induces the production of quantitatively and perhaps qualitatively different RNA when compared with RNA obtained from discs cultured without the hormone. The possible secondary structures of the RNA fractions are also discussed.
KeywordsGradient Centrifugation CsCl DEAE Ribonucleic Acid DEAE Cellulose
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