Emtage J S, Catlin G H, Carey N H
Nucleic Acids Res. 1979 Apr;6(4):1221-39. doi: 10.1093/nar/6.4.1221.
The polyadenylation of Fowl Plague Viral RNA and of Influenza A/Victoria Viral RNA using E. coli poly (A) polymerase and the subsequent reverse transcription of the polyadenylated species is reported. We have shown that all 8 genome fragments are adenylated and that an average of 25--30 adenylic acid residues per molecule is sufficient for maximal transcription with reverse transcriptase. The cDNA product is 95% sensitive to Sl-nuclease and hybridisation analysis against viral RNA reveals it to be a faithful copy of the RNA. Amongst the transcription products are long, discrete copies of genes 1--8, the lengths of which are comparable with those of the vRNA determined by electrophoresis on formamide acrylamide gels. These single-stranded cDNAs have been further transcribed to form double-stranded products with hair-pin structures at one end. Analysis of this material on native acrylamide gels revealed some DNA bands corresponding to the predicted sizes for genes 4--8.
报道了使用大肠杆菌聚（A）聚合酶对禽痘病毒RNA和甲型流感病毒/维多利亚病毒RNA进行聚腺苷酸化，以及随后对聚腺苷酸化产物进行逆转录的过程。我们已经表明，所有8个基因组片段都被腺苷酸化，并且每个分子平均25 - 30个腺苷酸残基足以实现逆转录酶的最大转录。cDNA产物对S1核酸酶的敏感性为95%，与病毒RNA的杂交分析表明它是RNA的忠实拷贝。转录产物中有基因1 - 8的长而离散的拷贝，其长度与通过在甲酰胺丙烯酰胺凝胶上电泳确定的vRNA长度相当。这些单链cDNA已进一步转录形成一端具有发夹结构的双链产物。在天然丙烯酰胺凝胶上对该材料的分析揭示了一些与基因4 - ⑧预测大小相对应的DNA条带。
