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首页> 外文期刊>Journal of Virology >Isolation of Defective Lysogens from Simian Virus 40-transformed Mouse Kidney Cultures
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Isolation of Defective Lysogens from Simian Virus 40-transformed Mouse Kidney Cultures

机译：硅藻病毒40转化小鼠肾脏培养物的缺陷溶血剂的分离

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Rescue of simian virus 40 (SV40) from hamster and murine cell lines transformed by nonirradiated or by ultraviolet (UV)-irradiated SV40 (10?3 to 10?5 survival) was studied. A combination of tests was employed to detect induction of SV40 synthesis: (i) co-cultivation with susceptible monkey kidney (CV-1) cells; (ii) treating mixtures of transformed and CV-1 cells with UV-irradiated Sendai virus (UV-Sendai) prior to co-cultivation; and (iii) plating untreated or UV-Sendai-treated mixtures of transformed and CV-1 cells with freshly trypsinized CV-1 cells. The first and second tests provided a measure of the total infectious SV40 yield per culture, and the third test provided a measure of the frequency of induction (fraction of transformed cells giving rise to infectious centers). With the combination of tests, SV40 was rescued in all trials from TSV-5 hamster cells, mKS-BU100 mouse cells, and from several lines of mouse kidney cells transformed by UV-irradiated SV40 (mKS-U lines). The frequency of induction was about 7 × 10?2 for TSV-5 cells, about 3 × 10?3 for mKS-BU100 cells, greater than 10?4 for the mKS-U lines which were “good” yielders, and about 10?5 to 10?4 for the mKS-U lines which were “average” yielders. SV40 of a plaque type different from parental virus was rescued from four of the mKS-U cell lines. Virus was also easily rescued from: (i) tumor cells produced from the mKS-A line of transformed mouse kidney cells; (ii) mouse kidney cells transformed by SV40 which had been rescued from mKS-BU100 cells; and (iii) tumor cells (HATS) which had been produced by inoculating newborn hamsters with SV40 rescued from mKS-BU100 cells. The frequency of induction of HATS cells was of the same order of magnitude as the frequency of induction of TSV-5 cells. In a study of the kinetics of virus induction, it was shown that SV40 could be detected 28, 40, and 48.5 hr after UV-Sendai treatment of mixtures of CV-1 and TSV-5, HATS, or mKS-BU100 cells, respectively. Although all of the mKS-U lines contained the SV40-specific tumor antigen, some were poor virus yielders (SV40 was recovered in less than 50% of the trials) and five lines were rare virus yielders (SV40 recovered only once in four or more trials). Forty-eight mKS-U lines were nonyielders; SV40 was never recovered by any test used thus far. UV-Sendai-treated mixtures of pairs of nonyielder mKS-U lines with CV-1 cells also did not yield infectious virus. Various factors affecting rescue have been discussed. The mKS-U lines which were poor virus yielders, rare yielders, or which never yielded virus have been classified tentatively as “defective lysogens” which contain mutational lesions at loci essential for detachment of SV40 from integration sites or for SV40 replication, or for both.
机译：从非辐射或通过紫外（UV）式SV40（10 3 / sop>至10 3 / sup>存活率，从仓鼠和鼠细胞系拯救羊毛病毒40（sv40）研究过。使用试验的组合来检测SV40合成的诱导：（i）用易感猴肾（CV-1）细胞共培养; （ii）在共培养之前将转化和CV-1细胞（UV-SENDAI）的转化和CV-1细胞的混合物治疗; （iii）用新胰蛋白酶化的CV-1细胞电镀未处理的或UV-Sendai处理的转化和CV-1细胞的混合物。第一和第二次测试提供了每种培养的总传染性SV40产量的量度，第三种测试提供了诱导频率的量度（转化细胞的一部分引起传染性中心）。随着测试的组合，SV40在来自TSV-5仓鼠细胞，MKS-Bu100小鼠细胞的所有试验中救出，以及由UV照射的SV40（MKS-U线）转化的几种小鼠肾细胞。诱导频率为TSV-5细胞的约7×10 2 ，约3×10 3 / sup>用于MKS-BU100电池，大于10 ？对于“良好”产量的MKS-U线，以及大约10 5 到10 ？4 的MKS-U线为“平均“屈服者。与父母病毒不同的斑块类型的SV40从4个MKS-U细胞系中救出。病毒也很容易从：（i）由MKS-A转化的小鼠肾细胞生产的肿瘤细胞; （ii）由SV40转化的小鼠肾细胞已从MKS-Bu100细胞中拯救; （III）通过接种来自MKS-BU100细胞的SV40的新生仓鼠生产的肿瘤细胞（帽）。帽子电池的诱导频率与诱导TSV-5细胞的频率相同的数量级。在病毒诱导的动力学的研究中，示出了SV40分别可以分别检测28,40和48.5小时的CV-1和TSV-5，帽子或MKS-BU100细胞的混合物后的28,40和48.5小时。虽然所有的MKS-U系列含有SV40特异性肿瘤抗原，但有些是差，病毒产量（SV40在少于50％的试验中回收），五条线稀有< / EM>病毒产量（SV40只在四次或更多试验中恢复一次）。四十八个MKS-U行是非rielders;迄今为止所使用的任何测试从未恢复SV40。 uv-sendai处理的与cv-1细胞对的对处理混合物也没有产生传染性病毒。已经讨论了影响救援的各种因素。病毒产量差，罕见的产量较差的MKS-U线或从未产生过的病毒已经暂时被分类为“有缺陷的溶血化”，其含有在基因座的突变病变，该突变病变是从整合位点分离或用于SV40复制的基因座，或者。

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