Munson L M, Reznikoff W S
Biochemistry. 1981 Apr 14;20(8):2081-5. doi: 10.1021/bi00511a003.
In vitro transcription assays have been used to study the rate of ribonucleic acid (RNA) synthesis from the Escherichia coli lactose promotor mutant lacL8UV5 contained on a 203-bp (base pair) restriction fragment. The half-life of long (63-base) RNA production from heparin-resistant RNA polymerase-promotor complexes was found to be related to the amount of oligonucleotides released during the initiation process (abortive initiation). Studies indicate that once a ternary complex between the promoter, RNA polymerase, and a newly synthesized RNA seven and nine nucleotides long is formed, abortive initiation is reduced and the rate of synthesis of long RNAs is increased. The promoter for the left inverted repeat of the transposable element Tn5 was also examined. It was observed to have a much slower rate of production of long RNAs, and it released oligonucleotides 4 times as often as the lactose promoter. The correlation between the amount of abortive initiation and the half-time of long RNA production is discussed.
体外转录分析已被用于研究从包含在一个203碱基对(bp)限制片段上的大肠杆菌乳糖启动子突变体lacL8UV5合成核糖核酸(RNA)的速率。发现从抗肝素RNA聚合酶 - 启动子复合物产生长(63碱基)RNA的半衰期与起始过程(流产起始)中释放的寡核苷酸量有关。研究表明,一旦在启动子、RNA聚合酶和新合成的7至9个核苷酸长的RNA之间形成三元复合物,流产起始就会减少,长RNA的合成速率就会增加。还研究了转座元件Tn5的左反向重复序列的启动子。观察到它产生长RNA的速率要慢得多,并且它释放寡核苷酸的频率是乳糖启动子的4倍。讨论了流产起始量与长RNA产生半衰期之间的相关性。