ISOLATION OF POLYSOMES FROM LARVAL AND ADULT DROSOPHILA-MELANOGASTER
ISOLATION OF POLYSOMES FROM LARVAL AND ADULT DROSOPHILA-MELANOGASTER
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DOI：
10.1016/0003-2697(81)90277-3
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发表时间：
1981-01-01
影响因子：
2.9
通讯作者：
SULLIVAN, DT
中科院分区：
文献类型：
--
作者：
BRADFORD, CT;SULLIVAN, DT
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A method was developed for the isolation of polysomes from adult D. melanogaster. The profiles are not degraded by endogenous nucleases at 4.degree. 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 of N-ethylmaleimide, cytidine 5''-monophosphoric acid, diethyl sulfate and polyvinylsulfate does not improve the profiles. The profiles are dependent on added Mg2+. Poly(A) 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.