Junghans R P, Duesberg P H, Knight C A
Proc Natl Acad Sci U S A. 1975 Dec;72(12):4895-9. doi: 10.1073/pnas.72.12.4895.
Varying the concentration of Triton X-100, a nonionic detergent used to promote the DNA polymerase activity of Rous sarcoma virus in an endogenous reaction, showed a very sharp peak at about 0.02% (vol/vol) for optimal DNA synthesis. The yield of DNA at this concentration of Triton exceeded yields obtained at concentrations above the optimum by a factor of 2-5 for the 90-min reaction. At optimal Triton concentration, about 1-7% of the DNA made in the absence of actinomycin and about 4-10% of the DNA made in the presence of actinomycin was 2.5 X 10(6) daltons or greater, as estimated by formamide polyacrylamide gel electrophoresis and by alkaline sucrose gradient sedimentation. No large DNA was obtained at higher than optimal Triton concentrations. The large DNA molecules were rendered totally resistant to single-strand specific nuclease S1 after hybridization to an excess of viral RNA. It was concluded that at optimal detergent concentration, the viral DNA polymerase can synthesize full-size DNA transcripts of viral RNA.
改变Triton X - 100（一种用于在内源反应中促进劳氏肉瘤病毒DNA聚合酶活性的非离子去污剂）的浓度，结果显示在大约0.02%（体积/体积）时DNA合成达到最优，出现一个非常尖锐的峰值。对于90分钟的反应，在此Triton浓度下的DNA产量比最优浓度以上的产量高出2至5倍。通过甲酰胺聚丙烯酰胺凝胶电泳和碱性蔗糖梯度沉降估计，在最优Triton浓度下，在无放线菌素时合成的DNA中约1 - 7%以及在有放线菌素时合成的DNA中约4 - 10%为2.5×10⁶道尔顿或更大。在高于最优Triton浓度时未获得大尺寸DNA。与过量病毒RNA杂交后，大尺寸DNA分子对单链特异性核酸酶S1完全具有抗性。得出的结论是，在最优去污剂浓度下，病毒DNA聚合酶能够合成病毒RNA的全长DNA转录本。
