Dubbs D R, Kit S, De Torres R A, Anken M
J Virol. 1967 Oct;1(5):968-79. doi: 10.1128/JVI.1.5.968-979.1967.
When simian virus 40 (SV40)-transformed mouse kidney cells (mKS) were grown in the presence of susceptible indicator cells, SV40 was readily recovered from: (i) 15 transformed cell lines, (ii) transformed cells subcultured 45 times over a 7-month period in medium containing antiviral serum and bromodeoxyuridine (dBU), (iii) 45 of 46 clonal lines isolated in the presence of antiviral serum, (iv) 19 of 19 secondary clones isolated from two clonal lines, and (v) dBU-resistant transformed cell lines. dBU-resistant SV40-transformed mouse kidney cell lines were selected and shown to contain the T antigen and to have normal levels of thymidylate kinase and deoxyribonucleic acid (DNA) polymerase, but to be deficient in thymidine (dT) kinase. Radioautographic and biochemical experiments demonstrated that very little (3)H-dT was incorporated into DNA of dBU-resistant cells during a 6-hr labeling period. After infection of dT kinase-deficient mKS cells with vaccinia virus, high levels of dT kinase were induced. The properties of SV40 recovered from dBU-sensitive and dBU-resistant cells were studied. SV40 recovered from transformed cells was shown to express in CV-1 cells at least six functions characteristic of parental virus: synthesis of capsid antigen, synthesis of T antigen, synthesis of viral DNA, induction of dT kinase, induction of DNA polymerase, and induction of host cell DNA synthesis. In addition, SV40 recovered from the transformed cells induced T antigen, dT kinase, deoxycytidylate deaminase, thymidylate kinase, and DNA polymerase in abortively infected mouse kidney cultures, and the virus was also capable of transforming primary cultures of mouse kidney cells.
当猿猴病毒40(SV40)转化的小鼠肾细胞(mKS)在易感指示细胞存在的情况下培养时,SV40很容易从以下细胞中回收:(i)15个转化细胞系;(ii)在含有抗病毒血清和溴脱氧尿苷(dBU)的培养基中在7个月内传代45次的转化细胞;(iii)在抗病毒血清存在下分离的46个克隆系中的45个;(iv)从两个克隆系中分离的19个二级克隆中的19个;以及(v)dBU抗性转化细胞系。选择了dBU抗性SV40转化的小鼠肾细胞系,结果显示其含有T抗原,胸苷酸激酶和脱氧核糖核酸(DNA)聚合酶水平正常,但胸苷(dT)激酶缺陷。放射自显影和生化实验表明,在6小时的标记期内,很少有(3)H-dT掺入dBU抗性细胞的DNA中。用痘苗病毒感染dT激酶缺陷的mKS细胞后,诱导出高水平的dT激酶。研究了从dBU敏感和dBU抗性细胞中回收的SV40的特性。从转化细胞中回收的SV40在CV-1细胞中显示出至少表达亲本病毒的六种特征性功能:衣壳抗原的合成、T抗原的合成、病毒DNA的合成、dT激酶的诱导、DNA聚合酶的诱导以及宿主细胞DNA合成的诱导。此外,从转化细胞中回收的SV40在流产感染的小鼠肾培养物中诱导T抗原、dT激酶、脱氧胞苷酸脱氨酶、胸苷酸激酶和DNA聚合酶,并且该病毒还能够转化小鼠肾细胞的原代培养物。