Isolation of specific messenger RNA by adsorption of polysomes to matrix-bound antibody
- 1 January 1977
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 4 (1) , 71-84
- https://doi.org/10.1093/nar/4.1.71
Abstract
A procedure is presented for the purification of specific mRNAs, which exploits the ability of antibodies prepared against a native protein to bind to the nascent polypeptide on the polysome. Rather than precipitating these soluble antibody-polysome complexes with anti-antibody, which canlead to nonspecific trapping of polysomes, we have linked the anti-antibody to an insoluble matrix. Thus, the antibody-polysome complex binds to the anti-antibody support and nonspecific polysomes can easily be removed by several washes. We have found para -aminobenzyl cellulose (PAB cellulose), to be a suitable matrix for this purpose. This support can bind large quantities of anti-antibody and it displayed no detectable nonspecific affinity for polysomes or RNA. Using this procedure, we have obtained an apparently homogeneous preparation of ovalbumin mRNA.Keywords
This publication has 24 references indexed in Scilit:
- Analysis of the complexity and diversity of mRNA from chicken liver and oviductCell, 1976
- Characterization of Polysomes from Xenopus Liver Synthesizing Vitellogenin and Translation of Vitellogenin and Albumin Messenger RNA's in vitroEuropean Journal of Biochemistry, 1976
- The isolation of chicken histone F2c (V) messenger RNA by immunoadsorption of F2c-synthesising polysomesBiochemical and Biophysical Research Communications, 1975
- Quantitation of parameters that determine the rate of ovalbumin synthesisCell, 1975
- Preparation and preliminary characterization of purified ovalbumin messenger RNA from the hen oviductBiochemistry, 1975
- Lineage-dependent transcription of globin genesCell, 1974
- Globin gene expression in cultured erythroleukemic cellsJournal of Molecular Biology, 1974
- Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undegraded polysomes and messenger ribonucleic acidBiochemistry, 1974
- Purification and Properties of Ovalbumin Messenger RNAEuropean Journal of Biochemistry, 1974
- Isolation of eukaryotic messenger RNA on cellulose and its translation in vitroBiochemical and Biophysical Research Communications, 1972