Roop D R, Tsai M J, O'Malley B W
Cell. 1980 Jan;19(1):63-8. doi: 10.1016/0092-8674(80)90388-8.
We present data which map the 5' and 3' ends of transcripts of the natural ovalbumin gene. First, hybridization of radiolabeled 5' and 3' fragments of the gene to oviduct nuclear RNA which had been electrophoresed in agarose gels and transferred to DBM paper localized the 5' and 3' ends of precursor molecules and demonstrated that the largest ovalbumin RNA molecules detectable by this method (approximately 7.8 kb) were similar in size to the ovalbumin natural gene. Second, hybridization of end-labeled probes to oviduct nuclear RNA followed by digestion with S1 nuclease and analysis on polyacrylamide gels mapped more precisely the 5' and 3' ends of the precursor molecules, and these termini were coincident with the beginning and end of the structural sequence of the natural gene. These results suggest that the 7.8 kb precursor is likely to be the primary transcript of the ovalbumin gene.
我们展示了定位天然卵清蛋白基因转录本5'端和3'端的数据。首先，将该基因经放射性标记的5'端和3'端片段与已在琼脂糖凝胶中进行电泳并转移至DBM纸上的输卵管核RNA进行杂交，从而定位前体分子的5'端和3'端，并证明通过该方法可检测到的最大卵清蛋白RNA分子（约7.8 kb）在大小上与天然卵清蛋白基因相似。其次，将末端标记的探针与输卵管核RNA杂交，随后用S1核酸酶消化并在聚丙烯酰胺凝胶上进行分析，更精确地定位了前体分子的5'端和3'端，并且这些末端与天然基因结构序列的起始和末尾一致。这些结果表明，7.8 kb的前体很可能是卵清蛋白基因的初级转录本。
