Ballario P, Buongiorno-Nardelli M, Carnevali F, Di Mauro E, Pedone F
Nucleic Acids Res. 1981 Aug 25;9(16):3959-78. doi: 10.1093/nar/9.16.3959.
The in vitro transcription properties of purified yeast RNA polymerase II have been analyzed on prokaryotic plasmids (pBR322 and pBR313) and chimaeric plasmids bearing yeast 2 micron sequences (BTYP 1, BTYH 2 and BTYH 3). Conditions for selective transcription of the 2 micron DNA sequences in chimaeric plasmids have been determined. pBR322 and pBR313 are not transcribed by the purified RNA polymerase II when not bearing eukaryotic inserts. We show that the agarose gel electrophoretic analysis of ternary transcription complexes allows the localization of nascent RNA chains. The RNA produced has been visualized by electron microscopy (nascent RNA hybridization loops) and by gel electrophoretic analysis. All the observed properties are shared by RNA polymerase II purified by a conventional method (1) and by a rapid alternative procedure described herein. The peculiar properties of a partially purified form of RNA polymerase II are reported.
已在原核质粒（pBR322和pBR313）以及带有酵母2微米序列的嵌合质粒（BTYP 1、BTYH 2和BTYH 3）上分析了纯化的酵母RNA聚合酶II的体外转录特性。已确定了嵌合质粒中2微米DNA序列选择性转录的条件。当不带有真核插入片段时，纯化的RNA聚合酶II不会转录pBR322和pBR313。我们表明，三元转录复合物的琼脂糖凝胶电泳分析可实现新生RNA链的定位。所产生的RNA已通过电子显微镜（新生RNA杂交环）和凝胶电泳分析进行了可视化。通过常规方法（1）纯化的RNA聚合酶II以及本文所述的快速替代方法纯化的RNA聚合酶II都具有所有观察到的特性。报道了一种部分纯化形式的RNA聚合酶II的特殊特性。
