Maden B E, Forbes J M, Stewart M A, Eason R
EMBO J. 1982;1(5):597-601. doi: 10.1002/j.1460-2075.1982.tb01214.x.
We have used two approaches to search for sequence variants in the 18S coding region of amplified ribosomal DNA (rDNA) from Xenopus laevis oocytes. First, using clones derived from amplified rDNA, we compared the equivalent of a complete 18S coding region from two clones and short regions from two other clones with the 18S sequence previously determined from a "reference" clone. The respective sequences in all the clones were identical. Secondly, we examined greater than 60% of the 18S sequence in "pooled 18S genes" in uncloned amplified rDNA. The predominant sequence corresponded to that in the reference clone and no heterogeneities were apparent. Since many chromosomal rDNA units contribute to rDNA amplification the findings indicate that 18S coding sequences in X. laevis are largely homogeneous. The previously established sequence is the predominant one, thus providing a reliable basis for studies on 18S rRNA. Sequencing gels on uncloned amplified rDNA confirmed the absence of methylated cytosine in this DNA. The 18S sequence lacks major open reading frames.
我们采用了两种方法来搜索非洲爪蟾卵母细胞中扩增核糖体DNA(rDNA)的18S编码区域的序列变体。首先,利用从扩增的rDNA中获得的克隆,我们将来自两个克隆的相当于完整18S编码区域以及另外两个克隆的短区域与先前从一个“参考”克隆中确定的18S序列进行了比较。所有克隆中的相应序列都是相同的。其次,我们检查了未克隆的扩增rDNA中“汇集的18S基因”中超过60%的18S序列。主要序列与参考克隆中的序列一致,没有明显的异质性。由于许多染色体rDNA单元参与rDNA扩增,这些发现表明非洲爪蟾的18S编码序列在很大程度上是同质的。先前确定的序列是主要序列,从而为18S rRNA的研究提供了可靠的基础。未克隆的扩增rDNA的测序凝胶证实了该DNA中不存在甲基化胞嘧啶。18S序列缺乏主要的开放阅读框。