Rapid purification of biologically active individual histone messenger RNAs by hybridization to cloned DNA linked to cellulose
We describe a rapid and simple method for the purification of biologically active messenger RNAs. The method allows the isolation in a few hours of specific mRNAs from either whole cell or polysomal RNA even if the RNA represents less than 1% of the starting molecules. We used, as a model, cloned se...
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| Veröffentlicht in: | Biochemistry (Easton) 1979-01, Vol.18 (1), p.208-213 |
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| container_title | Biochemistry (Easton) |
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| creator | Childs, Geoffrey Levy, Shoshana Kedes, Laurence H |
| description | We describe a rapid and simple method for the purification of biologically active messenger RNAs. The method allows the isolation in a few hours of specific mRNAs from either whole cell or polysomal RNA even if the RNA represents less than 1% of the starting molecules. We used, as a model, cloned sea urchin (Strongylocentrotus purpuratus) histone gene fragments linked to cellulose by the method of B. E. Noyes & G. R. Stark (1975) Cell 5, 301--310) as hybridization probes to isolate specific histone mRNAs from whole cell and polysomal RNA extracts. RNAs isolated in this manner maintain their biological activity, serving as templates for histone proteins in a wheat-germ, cell-free protein translation system. In addition, radiolabeled histone-specific RNA purified from cleavage stage sea urchin embryos, pulse labeled for short periods of time and analyzed on denaturing polyacrylamide gels, was the same size as mature histone mRNA's. |
| doi_str_mv | 10.1021/bi00568a032 |
| format | Article |
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The method allows the isolation in a few hours of specific mRNAs from either whole cell or polysomal RNA even if the RNA represents less than 1% of the starting molecules. We used, as a model, cloned sea urchin (Strongylocentrotus purpuratus) histone gene fragments linked to cellulose by the method of B. E. Noyes & G. R. Stark (1975) Cell 5, 301--310) as hybridization probes to isolate specific histone mRNAs from whole cell and polysomal RNA extracts. RNAs isolated in this manner maintain their biological activity, serving as templates for histone proteins in a wheat-germ, cell-free protein translation system. 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| ispartof | Biochemistry (Easton), 1979-01, Vol.18 (1), p.208-213 |
| issn | 0006-2960 1520-4995 |
| language | eng |
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| source | ACS Publications; MEDLINE |
| subjects | Animals Chromatography, Affinity DNA, Recombinant Embryo, Nonmammalian Escherichia coli - metabolism Female Histones - biosynthesis Molecular Weight Nucleic Acid Hybridization Plants - metabolism Plasmids Protein Biosynthesis RNA, Messenger - isolation & purification RNA, Messenger - metabolism RNA, Ribosomal - metabolism Sea Urchins - metabolism Triticum - metabolism |
| title | Rapid purification of biologically active individual histone messenger RNAs by hybridization to cloned DNA linked to cellulose |
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