Folk W R
J Virol. 1973 Mar;11(3):424-31. doi: 10.1128/JVI.11.3.424-431.1973.
BHK-21 cells were transformed with polyoma virus mutants Ts-a and Ts-25 by using a temperature shift from 31 to 39 C at 5 days after infection so that rescuable transformants could be isolated. Clones which yielded virus after fusion with mouse cells were scored and maintained at 39 C in the presence of antipolyoma virus antiserum. Generally, no infectious viral deoxyribonucleic acid (DNA) could be found in Hirt supernatant fractions of these lines when maintained at 39 C, but DNA-DNA reannealing measurements detected two to six viral genomes per diploid cell genome in the nuclear DNA. Fusion with permissive cells was not necessary to induce the synthesis of infectious virus; cell lines shifted to 31 C produce the equivalent of 100 viral genomes per cell after 5 days. In some cell lines up to 1% of the cells formed infectious centers upon a shift to 31 C, and 100% of the subclones of a line were inducible. Growth at 31 C selected for a noninducible population which was still transformed.
在感染后5天,通过将温度从31℃转变为39℃,用多瘤病毒突变体Ts-a和Ts-25转化BHK-21细胞,以便能够分离出可拯救的转化体。对与小鼠细胞融合后产生病毒的克隆进行评分,并在抗多瘤病毒抗血清存在的情况下于39℃进行维持培养。通常,当这些细胞系在39℃维持培养时,在Hirt上清液组分中找不到有感染性的病毒脱氧核糖核酸(DNA),但DNA-DNA复性测量在核DNA中检测到每个二倍体细胞基因组有2至6个病毒基因组。与允许性细胞融合并非诱导感染性病毒合成所必需;转移到31℃的细胞系在5天后每个细胞产生相当于100个病毒基因组的物质。在一些细胞系中,转移到31℃时高达1%的细胞形成了感染中心,并且一个细胞系的100%亚克隆都是可诱导的。在31℃生长选择出了一个仍处于转化状态的不可诱导群体。