Tsai S Y, Roop D R, Stumph W E, Tsai M J, O'Malley B W
Biochemistry. 1980 Apr 29;19(9):1755-61. doi: 10.1021/bi00550a005.
The transcription of DNA sequences flanking the 5' end and 3' end of the ovalbumin gene was examined. First, various restriction endonuclease fragments corresponding to the 5' and 3' regions of the gene isolated and used as hybridization probes to assay for the presence of transcripts corresponding to these different regions in the chick oviduct nuclear RNA. Very little, if any, of the transcripts corresponding to sequences flanking the 5' and 3' structural sequences of the ovalbumin gene was detected in the steady-state nuclear RNA. Second, RNA was pulse labeled either in isolated nuclei or in an oviduct tissue suspension system and hybridized to DNA filters containing purified fragments of various 5'- and 3'-flanking regions. Our results again demonstrated that RNA was not synthesized from the 5'- and 3'-flanking regions surrounding the gene. Taken together, these results are consistent with the postulate that flanking DNA sequences are not transcribed and that the largest RNA species detected in the nuclear RNA are the initial transcripts.
对卵清蛋白基因5'端和3'端侧翼的DNA序列转录情况进行了检测。首先,分离出与该基因5'和3'区域相对应的各种限制性内切酶片段,并将其用作杂交探针,以检测鸡输卵管核RNA中与这些不同区域相对应的转录本的存在情况。在稳态核RNA中,几乎检测不到(如果有的话)与卵清蛋白基因5'和3'结构序列侧翼序列相对应的转录本。其次,在分离的细胞核或输卵管组织悬浮系统中对RNA进行脉冲标记,并与含有各种5'和3'侧翼区域纯化片段的DNA滤膜杂交。我们的结果再次表明,基因周围的5'和3'侧翼区域未合成RNA。综上所述,这些结果与以下假设一致:侧翼DNA序列不被转录,并且在核RNA中检测到的最大RNA种类是初始转录本。