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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Kraus, J. P. Rosenberg, L. E. |
| Abstract | We have purified three low-abundance hepatic mRNAs to near homogeneity by polysome immunoadsorption. The mRNAs coding for the precursor of ornithine transcarbamoylase [carbamoylphosphate:L-ornithine carbamoyltransferase, EC 2.1.3.3], the precursor of the beta-subunit of propionyl-CoA carboxylase [propionyl-CoA:carbon dioxide ligase (ADP-forming), EC 6.4.1.3], and cystathionine beta-synthase [L-serine hydro-lyase (adding homocysteine), EC 4.2.1.22], representing approximately 0.20, 0.02, and 0.015% of total hepatic mRNA, respectively, were purified 450- to 6,300-fold. We used the following steps: interaction of rat liver polysomes with an IgG fraction of monospecific antisera raised against each polypeptide; immobilization of polysome-antibody complexes on a protein A-Sepharose column; removal of the bulk of polysomes by extensive washing; dissociation of ribosomal subunits and elution of specific mRNA with EDTA; and isolation of the eluted mRNA by chromatography on an oligo(dT)-cellulose column. It seems likely that this procedure will permit isolation of other low-abundance mRNAs and subsequent cloning of their respective cDNAs. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 13 |
| Volume Number | 79 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1982-10-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Carboxy-Lyases Genetics Cystathionine Beta-Synthase Hydro-Lyases Liver Enzymology Ornithine Carbamoyltransferase Polyribosomes Metabolism RNA, Messenger Isolation & Purification Animals Antigen-Antibody Complex Enzyme Precursors Immune Sera Methylmalonyl-CoA Decarboxylase Poly A Protein Biosynthesis RNA Research Support, U.S. Gov't, P.H.S. Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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