DesGroseillers L, Rassart E, Zollinger M, Jolicoeur P
J Virol. 1982 Apr;42(1):326-30. doi: 10.1128/JVI.42.1.326-330.1982.
We studied the synthesis of B-tropic murine leukemia viral DNA in vitro by detergent-disrupted virions. The reaction products (detected by the Southern transfer technique) included full-length, infectious, double-stranded DNA and several subgenomic fragments. Restriction endonuclease analysis and hybridization and specific probes revealed two classes of subgenomic fragments: some were derived from the right end of the genome, and some were derived from the left end. Most of the fragments harbored one long terminal repeat copy at their ends, suggesting that they were initiated correctly. S1 nuclease and restriction endonuclease treatments of these fragments indicated that a single-stranded gap was present near the first initiation site of plus strong-stop DNA. The treatments also suggested the presence of a second initiation site flanked by a single-stranded gap 0.9 kilobase pairs from the right end of the genome. Our data clearly show that plus-strand DNA is synthesized at both ends of the genome, by using plus strong stop as the first initiation site and additional initiation sites.
我们通过去污剂裂解的病毒粒子研究了体外B-嗜性鼠白血病病毒DNA的合成。反应产物（通过Southern印迹技术检测）包括全长、有感染性的双链DNA和几个亚基因组片段。限制性内切酶分析、杂交以及特异性探针显示存在两类亚基因组片段：一些来自基因组右端，一些来自左端。大多数片段在其末端含有一个长末端重复序列拷贝，表明它们起始正确。对这些片段进行S1核酸酶和限制性内切酶处理表明，在正链强终止DNA的第一个起始位点附近存在一个单链缺口。这些处理还表明，在距基因组右端0.9千碱基对处有一个被单链缺口侧翼的第二个起始位点。我们的数据清楚地表明，正链DNA是在基因组两端合成的，以正链强终止作为第一个起始位点以及其他起始位点。
