Bollen G H, Mager W H, Jenneskens L W, Planta R J
Eur J Biochem. 1980 Mar;105(1):75-80. doi: 10.1111/j.1432-1033.1980.tb04475.x.
In order to identify and to study the ribosomal protein genes in yeast we have tried to purify the mRNA coding for ribosomal proteins. Poly(A)-containing RNA from the yeast Saccharomyces carlsbergensis was fractionated according to size using preparative sucrose gradient centrifugation. The various (size) fractions were translated in vitro in a wheat germ cell-free system. The products were analysed by sodium dodecylsulfate/polyacrylamide gradient gel electrophoresis as well as by acetic acid/urea gel electrophoresis. It was found that an mRNA fraction of about 9 S directs the synthesis in vitro of proteins that have properties characteristic of ribosomal proteins, i.e. they are both small and basic. The ribosomal nature of these proteins was further established by two-dimensional gel electrophoresis. This small-size mRNA fraction can be used as a probe for the identification of ribosomal protein genes in recombinant DNA molecules.
为了鉴定和研究酵母中的核糖体蛋白基因，我们尝试纯化编码核糖体蛋白的mRNA。使用制备型蔗糖梯度离心法，根据大小对来自卡尔酵母（Saccharomyces carlsbergensis）的含聚腺苷酸（Poly(A)）RNA进行分级分离。将各个（大小）级分在小麦胚芽无细胞体系中进行体外翻译。通过十二烷基硫酸钠/聚丙烯酰胺梯度凝胶电泳以及乙酸/尿素凝胶电泳对产物进行分析。发现约9S的一个mRNA级分在体外指导合成具有核糖体蛋白特性的蛋白质，即它们既小又呈碱性。通过二维凝胶电泳进一步确定了这些蛋白质的核糖体性质。这个小尺寸的mRNA级分可用作探针，用于鉴定重组DNA分子中的核糖体蛋白基因。
