Yukawa Y, Sugita M, Sugiura M
Center for Gene Research, Nagoya University, Japan.
Plant J. 1997 Oct;12(4):965-70. doi: 10.1046/j.1365-313x.1997.12040965.x.
A nuclear extract derived from tobacco cultured BY-2 cells supports RNA polymerase III-dependent transcription of Arabidopsis tRNA(Ser) genes. Primer extension analysis indicated that the transcription starts at 6 bp upstream from the 5' end of tRNA coding region. Procedures for nuclear extraction and in vitro reaction conditions have been optimized for tRNA transcription, which allows direct detection of de novo synthesized tRNA by gel electrophoresis. This improved in vitro system yields a mature-sized tRNA of 85 nucleotides from the Arabidopsis tRNA(Ser) gene, indicating that efficient processing of the pre-tRNA also occurs in the tobacco nuclear extract.
源自烟草培养的BY-2细胞的核提取物支持拟南芥tRNA(Ser)基因的RNA聚合酶III依赖性转录。引物延伸分析表明转录起始于tRNA编码区5'端上游6个碱基处。已针对tRNA转录优化了核提取程序和体外反应条件，这使得通过凝胶电泳能够直接检测从头合成的tRNA。这个改进的体外系统从拟南芥tRNA(Ser)基因产生了一个85个核苷酸的成熟大小的tRNA，表明前体tRNA的有效加工也发生在烟草核提取物中。
