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 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.
D000818:Animals; D002846:Chromatography, Affinity; D004274:DNA, Recombinant; D004625:Embryo, Nonmammalian; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D006657:Histones / Q000096:biosynthesis*; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D010944:Plants / Q000378:metabolism; D010957:Plasmids; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism; D012617:Sea Urchins / Q000378:metabolism; D014908:Triticum / Q000378:metabolism
D004274:DNA, Recombinant; D006657:Histones; D012333:RNA, Messenger; D012335:RNA, Ribosomal