Gubler U, Wyler T, Seebeck T, Braun R
Nucleic Acids Res. 1980 Jun 25;8(12):2647-64. doi: 10.1093/nar/8.12.2647.
The processing of intermediates of Physarum rRNA has been investigated using two cloned rDNA restriction fragments as sequence specific probes. The size of the largest stable pre-rRNA detected is 11.8 Kb, as determined by denaturing gel electrophoresis. R-loop analysis revealed that this pre-rRNA fraction mainly consists of transcripts which do not contain the two insertion sequences present in the 26 S gene. However, a small number of molecules are found which still contain one or the other, or both, of these introns. These observations suggest that the two introns are transcribed but spliced out in a random order before RNA synthesis has reached the 3-end of the primary transcript. Transcription starts at about 17.7 Kb and stops at 4.4 Kb as measured from the ends of the linear palindromic rDNA molecule. Besides the 11.8 Kb transcript several further ribosomal RNA precursors have been identified, ranging in size from 2.5 to 8.8 Kb. In addition, the genes for 5.8 S RNA have been located by R-loop mapping.
利用两个克隆的rDNA限制性片段作为序列特异性探针,对绒泡菌rRNA中间体的加工过程进行了研究。通过变性凝胶电泳测定,检测到的最大稳定前体rRNA的大小为11.8 kb。R环分析表明,该前体rRNA部分主要由不包含26S基因中存在的两个插入序列的转录本组成。然而,发现少量分子仍然包含这些内含子中的一个或另一个,或两者都有。这些观察结果表明,这两个内含子被转录,但在RNA合成到达初级转录本的3'末端之前以随机顺序被剪接掉。从线性回文rDNA分子的末端测量,转录起始于约17.7 kb处,终止于4.4 kb处。除了11.8 kb的转录本外,还鉴定出了几种大小在2.5至8.8 kb之间的其他核糖体RNA前体。此外,通过R环作图确定了5.8S RNA的基因位置。