T7 RNA polymerase transcription of Escherichia coli isoacceptors tRNALeu
T7 RNA polymerase transcription of Escherichia coli isoacceptors tRNALeu
复制标题
DOI：
10.1007/bf02880055
复制
发表时间：
1999-04-01
期刊：
影响因子：
--
通讯作者：
Wang, YL
中科院分区：
文献类型：
--
作者：
Li, Y;Chen, JF;Wang, YL
化学合成T7启动子和编码tRNA的基因（Leu），克隆到质粒pUC 19中，用纯化的T7 RNA聚合酶体外转录。发现亚精胺对T7转录有负面影响。在最佳反应条件下，转录的tRNA量约为模板DNA的250倍。未修饰的tRNA（1）（Leu）和tRNA（2）（Leu）具有相似的亮氨酸接受能力，但仅为天然修饰的tRNA的四分之一。这一结果表明，tRNA的修饰核苷酸（Leu）在其氨酰化过程中发挥了一定的作用，但不是关键作用。
The T7 promoter and the gene encoding tRNA(Leu) were synthesized chemically and cloned into plasmid pUC19, where the gene was transcribed in vitro with T7 RNA polymerase purified. It was found that spermidine had a negative effect on T7 transcription. The amount of transcribed tRNA was about 250 times that of the template DNA under optimal reaction conditions. The unmodified tRNA(1)(Leu) and tRNA(2)(Leu) had similar leucine accepting ability, which however was only one fourth that of the native modified ones. This result indicated that the modified nucleotides of tRNA(Leu) played some though not key roles in its aminoacylation.