Kit S, Brown M
Division of Biochemical Virology, Baylor College of Medicine, Houston, Texas 77025.
J Virol. 1969 Sep;4(3):226-30. doi: 10.1128/JVI.4.3.226-230.1969.
The relation between simian virus 40 (SV40) input multiplicity during transformation of primary mouse kidney cultures and the subsequent rescue of SV40 from clonal lines of transformed cells has been studied. Primary mouse kidney cultures were transformed with unirradiated SV40 at input multiplicities varying from 0.06 to 200 plaque-forming units (PFU) /cell or with SV40 irradiated with ultraviolet (UV) light to a survival of 0.04 to 0.01. All of the transformed lines contained the intranuclear SV40 T antigen, but cell-free extracts prepared from the transformed cell lines failed to yield infectious virus when assayed on monkey kidney cell (CV-1) monolayers. After fusion with susceptible CV-1 cells induced by UV-inactivated Sendai, all of the lines transformed by unirradiated virus yielded infectious SV40. The frequency of induction and the incidence of successful trials did not depend on the multiplicity of infection. "Good" yielders were obtained from mouse kidney cells transformed at the low input multiplicity of 0.06 PFU /cell. In contrast, only 4 of 12 clonal lines transformed at moderately low input multiplicity, and none of the lines transformed at very low input multiplicity with UV-irradiated virus yielded infectious SV40. The four positive lines have been classified as "poor" or "rare" yielders.
研究了原代小鼠肾细胞培养物转化过程中猿猴病毒40(SV40)的输入复数与随后从转化细胞克隆系中拯救SV40之间的关系。用未辐照的SV40以0.06至200蚀斑形成单位(PFU)/细胞的不同输入复数或用紫外线(UV)照射至存活率为0.04至0.01的SV40转化原代小鼠肾细胞培养物。所有转化系均含有核内SV40 T抗原,但从转化细胞系制备的无细胞提取物在猴肾细胞(CV-1)单层上检测时未能产生感染性病毒。在用紫外线灭活的仙台病毒诱导与易感CV-1细胞融合后,所有用未辐照病毒转化的系均产生了感染性SV40。诱导频率和成功试验的发生率不取决于感染复数。“高产”系是从以0.06 PFU/细胞的低输入复数转化的小鼠肾细胞中获得的。相比之下,在中等低输入复数下转化的12个克隆系中只有4个,而在用紫外线辐照病毒以非常低的输入复数转化的系中没有一个产生感染性SV40。这四个阳性系已被归类为“低产”或“罕见”产毒株。