Leader D J, Clark G P, Brown J W
Department of Cell and Molecular Genetics, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK.
Biochim Biophys Acta. 1998 May 11;1397(3):325-30. doi: 10.1016/s0167-4781(98)00025-6.
Northern analyses of U14snoRNAs in different plant species showed the expected hybridising band of approximately 120 nt in monocotyledonous and dicotyledonous angiosperms. In the lower plant, Bird's nest fern (Asplenium nidus), U14s were larger and three hybridising RNAs of approximately 190, 210 and 250 nt were observed. RT-PCR cloning of all three size variants using primers to the conserved 5' and 3' ends of higher plant U14snoRNAs showed large insertions in one of the plant-specific regions corresponding in position to the yeast U14-specific Y-domain. The insertions are pyrimidine-rich in their 5' halves and purine-rich in their 3' halves and are likely to be sequestered in stem structures consistent with the proposed model of U14snoRNA secondary structure. The 5' flanking regions of one of the fern U14 variants was generated by PCR and lacked classical plant snRNA promoter elements.
对不同植物物种中U14小核仁RNA(snoRNA)的Northern印迹分析表明,在单子叶和双子叶被子植物中出现了预期的约120个核苷酸的杂交条带。在低等植物鸟巢蕨(Asplenium nidus)中,U14更大,观察到三种杂交RNA,大小约为190、210和250个核苷酸。使用针对高等植物U14 snoRNA保守5'和3'末端的引物对所有三种大小变体进行逆转录聚合酶链反应(RT-PCR)克隆,结果显示在与酵母U14特异性Y结构域位置相对应的一个植物特异性区域中有大的插入片段。这些插入片段在其5'半段富含嘧啶,在其3'半段富含嘌呤,并且很可能被隔离在与所提出的U14 snoRNA二级结构模型一致的茎结构中。通过聚合酶链反应(PCR)产生了一种蕨类植物U14变体的5'侧翼区域,该区域缺乏经典的植物小核RNA(snRNA)启动子元件。