Klekamp M S, Weil P A
J Biol Chem. 1982 Jul 25;257(14):8432-41.
Cell-free extracts prepared from whole yeast cells carry out selective and accurate transcription, in vitro, of purified yeast class III genes. Both 5 S rRNA and tRNA genes are specifically transcribed by DNA-dependent RNA polymerase III present in these whole cell extracts. These extracts also appear to carry out nucleolytic processing of the in vitro synthesized transcripts. Optimal conditions for specific class III gene transcription in vitro are defined. Initial fractionation of the yeast extract has indicated that multiple chromatographically separable factors (fractions) are required, in addition to RNA polymerase III, for specific in vitro transcription of class III genes.
从全酵母细胞制备的无细胞提取物在体外对纯化的酵母III类基因进行选择性和精确转录。5S rRNA基因和tRNA基因均由这些全细胞提取物中存在的DNA依赖性RNA聚合酶III特异性转录。这些提取物似乎还对体外合成的转录本进行核酸酶加工。确定了体外特异性III类基因转录的最佳条件。酵母提取物的初步分级分离表明,除RNA聚合酶III外,III类基因的特异性体外转录还需要多个可通过色谱分离的因子(组分)。