Grummt I, Skinner J A
Proc Natl Acad Sci U S A. 1985 Feb;82(3):722-6. doi: 10.1073/pnas.82.3.722.
The activity of the mouse ribosomal promoter was examined after fusion to the gene coding for chloramphenicol acetyltransferase (CAT) and transfection into mouse cells. Very little CAT enzyme but high levels of CAT-specific RNA correctly initiated at the ribosomal DNA start site were synthesized. The amount of specific transcripts was neither influenced by long stretches of upstream spacer sequences nor by the insertion of the Moloney murine sarcoma virus enhancer. The deletion mutant pMr delta-39, which has been shown to be fully active in vitro, exhibited a 90% decrease in template activity in vivo. A mutant in which 22 base pairs of ribosomal DNA (between positions -35 and -14) were substituted by foreign DNA sequences proved transcriptionally inactive. The fusion genes were only transcribed in mouse cells, indicating that species-specific transcription factors are involved in ribosomal promoter recognition.
将氯霉素乙酰转移酶(CAT)编码基因与小鼠核糖体启动子融合,并转染到小鼠细胞后,对其活性进行了检测。合成的CAT酶很少,但在核糖体DNA起始位点正确起始的CAT特异性RNA水平很高。特异性转录本的量既不受上游间隔序列长片段的影响,也不受莫洛尼鼠肉瘤病毒增强子插入的影响。已证明在体外完全有活性的缺失突变体pMr delta-39,在体内模板活性降低了90%。一个核糖体DNA的22个碱基对(在-35和-14位之间)被外源DNA序列取代的突变体被证明转录无活性。融合基因仅在小鼠细胞中被转录,这表明物种特异性转录因子参与核糖体启动子的识别。