Hanano S, Sugita M, Sugiura M
Center for Gene Research, Nagoya University, Japan.
DNA Res. 1996 Apr 30;3(2):65-71. doi: 10.1093/dnares/3.2.65.
We have previously characterized a tobacco cDNA encoding a novel type RNA-binding protein (RZ-1), which contains a zinc finger motif in addition to a consensus sequence-type RNA-binding domain and is localized in the nucleus. Here we isolated its genomic clone from a Nicotiana sylvestris genomic library. Southern blot analysis suggested that RZ-1 is coded for by a single locus per haploid genome. Comparison of the cDNA and genomic sequences indicated that the RZ-1 gene contains two introns, one in the coding region and another in the 3'-untranslated region. RT-PCR and ribonuclease protection analyses showed that splicing of RZ-1 pre-mRNA occurs efficiently. The RZ-1 protein is actively synthesized in rapidly dividing tobacco cells, as demonstrated by immunoblot analysis.
我们之前鉴定了一个编码新型RNA结合蛋白(RZ-1)的烟草cDNA,该蛋白除了共有序列型RNA结合域外还含有一个锌指基序,并且定位于细胞核中。在此,我们从野生烟草基因组文库中分离出了它的基因组克隆。Southern印迹分析表明,单倍体基因组中每个位点编码一个RZ-1。cDNA与基因组序列的比较表明,RZ-1基因含有两个内含子,一个在编码区,另一个在3'非翻译区。RT-PCR和核糖核酸酶保护分析表明,RZ-1前体mRNA的剪接有效发生。免疫印迹分析表明,RZ-1蛋白在快速分裂的烟草细胞中被活跃合成。