 A method for isolation of intact, translationally active ribonucleic acid; ; et al in DNA (1983), 2(4), 329-35 A method for isolation of large, translationally active RNA species is presented. The procedure involves homogenization of cells or tissues in 5 M guanidine monothiocyanate followed by direct ... [more ▼] A method for isolation of large, translationally active RNA species is presented. The procedure involves homogenization of cells or tissues in 5 M guanidine monothiocyanate followed by direct precipitation of RNA from the guanidinium by 4 M LiCl. Modifications are described for use with tissue culture cells, yeast, tissues, or isolated nuclei. The advantages of the procedure include speed, simplicity, avoidance of an ultracentrifugation, and its applicability to large numbers of small samples. The procedure yields large mRNA precursors up to 10 kb and mRNA species which translate very well. However, small (less than 300 nucleotides) RNA species are recovered with a poor yield. [less ▲] Detailed reference viewed: 87 (0 ULg)Cell-free synthesis of acetylcholine receptor polypeptides; ; Martial, Joseph  et alin Science (1980), 209(4457), 695-7 Messenger RNA coding for acetylcholine receptor peptides has been identified. This polyadenylate [poly(A)+]RNA from Torpedo californica directs, in a cell-free system, the synthesis of peptides 60,000, 51 ... [more ▼] Messenger RNA coding for acetylcholine receptor peptides has been identified. This polyadenylate [poly(A)+]RNA from Torpedo californica directs, in a cell-free system, the synthesis of peptides 60,000, 51,000, 49,000 41,000, and 35,000 daltons which account for approximately 2 percent of the total synthesized proteins. The results suggest that several different messenger RNA's code for the receptor subunits. These proteins react specifically to antiserum to native acteylcholine receptor, suggesting that the primary translational product has conformational features similar to the native receptor. Further, the results support the idea that there is post-translational modification of receptor subunits as the molecular weights of the cell-free synthesized proteins differ from those of purified receptor subunits. [less ▲] Detailed reference viewed: 12 (3 ULg) |
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