Majumder H K, Maitra U, Rosenberg M
Proc Natl Acad Sci U S A. 1979 Oct;76(10):5110-3. doi: 10.1073/pnas.76.10.5110.
RNA was synthesized in vitro from a T3 DNA template by T3 RNA polymerase and subsequently separated into seven discrete size classes (molecular weights ranging between 0.21 x 10(6) and 6.2 x 10(6)) by electrophoresis in polyacrylamide slab gels. RNase T1-generated 3'-terminal oligonucleotide fragments were then selectively isolated from either the unfractionated total RNA or the gel-purified specific transcripts by chromatography on columns of dihydroxyboryl-cellulose. Sequence analysis of these oligonucleotide products indicated that the unfractionated transcripts as well as all the individual major RNA species examined had a unique sequence, (Gp)UpUpUpUpUpGOH, at their 3' termini. The specificity of this sequence, as well as the total lack of any sequence heterogeneity at the ends of these transcripts, indicates a high degree of specificity of termination during transcription in this system.
RNA 通过 T3 RNA 聚合酶从 T3 DNA 模板体外合成，随后通过在聚丙烯酰胺平板凝胶中电泳分离成七个离散的大小类别（分子量在 0.21×10⁶ 至 6.2×10⁶ 之间）。然后通过在二羟基硼基纤维素柱上进行色谱法，从未分级的总 RNA 或凝胶纯化的特定转录本中选择性分离出 RNase T1 产生的 3'-末端寡核苷酸片段。对这些寡核苷酸产物的序列分析表明，未分级的转录本以及所有检测的单个主要 RNA 物种在其 3' 末端都有一个独特的序列 (Gp)UpUpUpUpUpGOH。该序列的特异性以及这些转录本末端完全缺乏任何序列异质性，表明该系统转录过程中终止具有高度特异性。
