Isolation of polysomes from larval and adult Drosophila melanogaster

✍ Scribed by Carol T. Bradford; David T. Sullivan

• Book ID: 102985693
• Publisher: Elsevier Science
• Year: 1981
• Tongue: English
• Weight: 591 KB
• Volume: 112
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(81)90277-3

✦ Synopsis

The profiles are not degraded by endogenous nucleases at 4"C, but are sensitive to added ribonuclease A. Cycloheximide added to the preparation does not alter the profile, indicating that chain elongation leading to loss of polysomes is not occurring under these conditions. Resuspension of the polysome pellets in high-salt buffer leads to dissociation of inactive ribosomes which is reversible. The addition ofN-ethylmaleimide, cytidine Y-monophosphoric acid, diethyl sulfate, and polyvinylsulfate does not improve the protiles. The profiles are dependent on added magnesium ion. Polyadenylated RNA was extracted from the polysomes using the guanidinium chloride method and oligo(dT)-cellulose chromatography. The RNA was translated in a rabbit reticulocyte lysate cell-free system and the translation products were analyzed by polyacrylamide gel electrophoresis.