Edwards J C, Levens D, Rabinowitz M
Cell. 1982 Dec;31(2 Pt 1):337-46. doi: 10.1016/0092-8674(82)90127-1.
We have developed an in vitro transcription system for yeast mitochondrial rRNA genes. Using highly purified yeast mitochondrial RNA polymerase and bacterial plasmids carrying DNA segments containing the mitochondrial rRNA sites of transcriptional initiation, we have been able to demonstrate correct initiation of transcription in vitro. By directly sequencing the transcription products, we show that transcription in vitro of both the 14S and 21S rRNAs is initiated at precisely the same site as it is in vivo. Transcription of the rRNA genes is highly sensitive to ionic strength and RNA polymerase concentration. Additional factors or modified conditions may be necessary to permit accurate transcription of mitochondrial protein genes.
我们已经开发出一种用于酵母线粒体rRNA基因的体外转录系统。利用高度纯化的酵母线粒体RNA聚合酶以及携带包含线粒体rRNA转录起始位点DNA片段的细菌质粒，我们已经能够证明体外转录的正确起始。通过直接对转录产物进行测序，我们表明14S和21S rRNA的体外转录起始位点与体内完全相同。rRNA基因的转录对离子强度和RNA聚合酶浓度高度敏感。可能需要其他因子或改良条件才能实现线粒体蛋白质基因的准确转录。
