Staub J M, Castora F J
Waksman Institute, Rutgers University, Piscataway, NJ 08855.
Biochem Biophys Res Commun. 1993 Apr 30;192(2):616-26. doi: 10.1006/bbrc.1993.1460.
We have used a prokaryotic terminator identification vector, pDR721, to isolate regions from rat mitochondrial DNA (mtDNA) that can act as transcription terminators in vivo. Three independent fragments having terminator capability have been mapped to three general regions of the mitochondrial genome. Two terminators, pRMT1 and pRMT3, are found within and around the D-loop and cytochrome b gene, respectively, while the third, pRMT5, is located at the 3'-end of the 16S ribosomal RNA gene. After subcloning into host cells which carried temperature sensitive mutations in the termination factor, rho protein, galactokinase assays at the permissive and non-permissive temperatures suggested that pRMT3 acted as a rho-independent termination element while the other two, pRMT1 and pRMT5, were dependent on rho protein (or a rho-like protein) for efficient transcription termination.
我们使用了一种原核生物终止子鉴定载体pDR721,从大鼠线粒体DNA(mtDNA)中分离出在体内可作为转录终止子的区域。具有终止子能力的三个独立片段已被定位到线粒体基因组的三个大致区域。两个终止子pRMT1和pRMT3分别位于D环和细胞色素b基因内部及周围,而第三个终止子pRMT5位于16S核糖体RNA基因的3'端。将这些片段亚克隆到携带终止因子rho蛋白温度敏感突变的宿主细胞后,在允许温度和非允许温度下进行的半乳糖激酶测定表明,pRMT3作为一种不依赖rho的终止元件起作用,而另外两个pRMT1和pRMT5则依赖rho蛋白(或类似rho的蛋白)进行有效的转录终止。