Ribosomal subunit antiassociation activity in rabbit reticulocyte lysates. Evidence for a low molecular weight ribosomal subunit antiassociation protein factor (Mr = 25,000).
1984; Elsevier BV; Volume: 259; Issue: 19 Linguagem: Inglês
10.1016/s0021-9258(20)71300-1
ISSN1083-351X
AutoresPradip Raychaudhuri, E A Stringer, David M. Valenzuela, Umadas Maitra,
Tópico(s)Transgenic Plants and Applications
ResumoRabbit Reticulocyte RibosomalSubunit Antiassociation Factor 11931suspended in 20 mM Tris-HC1, pH 7.5, 100 mM KCl, 5 mM MgC12, 1 mM dithiothreitol, and 50% glycerol and stored at -80 "C at a concentration of 400 AZM) unit/ml.One A m unit of 40 S, 60 S, and 80 S ribosomes is equivalent to 65, 29, and 21 pmol, respectively.Rabbit reticulocyte lysates were either prepared from New Zealand Black rabbits treated with acetylphenylhydrazine according to the method of Ranu and London (15) or obtained from Green Hectares Co., Oregon, WI.Assay of Ribosomal Subunit Antiassociation Factor Activity-The assay method is similar to that described previously (11) and is based on the principle that the antiassociation factor prevents the spontaneous reassociation of eukaryotic ribosomal subunits when the M$+ concentration is increased from 1 to 5 mM.Reaction mixtures (75 pl) contained 20 mM Tris-HC1, pH 7.5, 100 mM KC1, 1 mM MgC12, 1 mM dithiothreitol, and 0.18 AZM) unit of either A. salina or rabbit reticulocyte 80 S ribosomes or 40 S and 60 S ribosomal subunits in the amount indicated in the figures as well
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