Richter K, Ammerer G, Hartter E, Ruis H
J Biol Chem. 1980 Sep 10;255(17):8019-22.
Total RNA was isolated from mutants of Saccharomyces cerevisiae that lack active delta-aminolevulinate synthase and are therefore defective in heme biosynthesis. The RNAs were translated in the cell-free protein synthesis system from wheat germ, and the catalase T synthesized was isolated by immunoadsorption and polyacrylamide gel electrophoresis in the presence of dodecyl sulfate. Little or no catalase T product was detected with RNA from mutant cells grown in the absence of a heme precursor. As judged from in vitro translational capacity, RNA fractions from mutant cells grown in the presence of delta-aminolevulinate contained at least 10 times more catalase T mRNA than RNA from unsupplemented cells.
从缺乏活性δ-氨基乙酰丙酸合酶、因此血红素生物合成有缺陷的酿酒酵母突变体中分离出总RNA。这些RNA在来自小麦胚芽的无细胞蛋白质合成系统中进行翻译，通过免疫吸附和在十二烷基硫酸钠存在下的聚丙烯酰胺凝胶电泳分离合成的过氧化氢酶T。在用缺乏血红素前体的培养基培养的突变体细胞的RNA中，几乎检测不到或没有检测到过氧化氢酶T产物。根据体外翻译能力判断，在δ-氨基乙酰丙酸存在下培养的突变体细胞的RNA组分中过氧化氢酶T mRNA的含量比未添加该物质的细胞的RNA至少多10倍。
