Guerineau F, Waugh R
Scottish Crop Research Institute, Invergowrie, Dundee, UK.
Plant Mol Biol. 1993 Aug;22(5):807-18. doi: 10.1007/BF00027367.
Using the inverse polymerase chain reaction (IPCR), 19 U6snRNA gene promoters were isolated from the potato genome. Analysis of their nucleotide sequences revealed the existence of two subfamilies. Promoters from class 1 harbour the typical sequence elements required for plant snRNA gene transcription whereas those from class 2 do not have a TATA box. Three promoters were fused to a modified U6snRNA-coding sequence to allow their activity to be monitored in tobacco protoplasts. Two of the promoters, one from either class, were found to be active. Comparison of potato U6snRNA gene promoter sequences with those found in other plant species showed various degrees of homology. In addition, the entire nucleotide sequences of seven potato U6snRNA genes and one pseudogene were determined. The overall frequency of nucleotide changes after PCR was found to be 1.15 x 10(-3). The mutations appeared to be clustered in a distinct area and were all A-to-G/T-to-C substitutions.
利用反向聚合酶链反应(IPCR),从马铃薯基因组中分离出19个U6snRNA基因启动子。对其核苷酸序列的分析揭示了两个亚家族的存在。1类启动子含有植物snRNA基因转录所需的典型序列元件,而2类启动子没有TATA框。将三个启动子与修饰的U6snRNA编码序列融合,以便在烟草原生质体中监测它们的活性。发现其中两个启动子有活性,每个类各一个。将马铃薯U6snRNA基因启动子序列与其他植物物种中的启动子序列进行比较,发现它们具有不同程度的同源性。此外,还测定了7个马铃薯U6snRNA基因和1个假基因的完整核苷酸序列。发现PCR后核苷酸变化的总体频率为1.15×10⁻³。这些突变似乎聚集在一个特定区域,并且都是A到G/T到C的替换。