Craig E A, Raskas H J
Cell. 1976 Jun;8(2):205-13. doi: 10.1016/0092-8674(76)90004-0.
Nuclear viral RNAs synthesized early in productive infection with adenovirus 2 were analyzed by hybridization to specific viral DNA fragments. Radioactive RNAs were subjected to electrophoresis in polyacrylamide gels containing 98% formamide, and the fractionated RNA was hybridized to specific DNA fragments generated by cleavage with endo R-Eco Ri or endo R-Sma l. The viral genes expressed early in infection are located in four different segments of the genome. When nuclear RNA was hybridized to DNA fragments representing each of these segments, discrete RNA size classes were detected. For each of these four regions of the genome, some of the discrete nuclear RNAs were larger than the cytoplasmic mRNAs. As calculated from electrophoretic mobility, the molecular weight of these nuclear RNA size classes ranged from 15% to several fold greater than that of the corresponding cytoplasmic mRNAs. Hybridization-inhibition experiments were performed to analyze the sequences present in one of these larger nuclear transcripts, a 22S RNA transcribed from the left-hand end of the genome. This 22S nuclear RNA was transcribed from the same strand as the viral mRNA, for it contained cytoplasmic sequences; it also contained sequences restricted to the nucleus. The larger nuclear RNAs may serve as precursors to the cytoplasmic viral mRNAs synthesized early in infection.
通过与特定病毒DNA片段杂交,分析了腺病毒2生产性感染早期合成的核病毒RNA。将放射性RNA在含有98%甲酰胺的聚丙烯酰胺凝胶中进行电泳,然后将分级分离的RNA与用内切酶R-EcoRⅠ或内切酶R-SmaⅠ切割产生的特定DNA片段杂交。感染早期表达的病毒基因位于基因组的四个不同区段。当核RNA与代表这些区段中每一个的DNA片段杂交时,检测到离散的RNA大小类别。对于基因组的这四个区域中的每一个,一些离散的核RNA比细胞质mRNA大。根据电泳迁移率计算,这些核RNA大小类别的分子量比相应细胞质mRNA的分子量高15%到几倍。进行杂交抑制实验以分析这些较大核转录本之一(从基因组左端转录的22S RNA)中存在的序列。这种22S核RNA与病毒mRNA转录自同一条链,因为它包含细胞质序列;它还包含仅限于细胞核的序列。较大的核RNA可能是感染早期合成的细胞质病毒mRNA的前体。