Allen G P, McGowan J J, Gentry G A, Randall C C
J Virol. 1978 Oct;28(1):361-7. doi: 10.1128/JVI.28.1.361-367.1978.
A line of 3T3 mouse cells lacking deoxythymidine kinase (dTK-) was stably transformed to the dTK+ phenotype after exposure to UV-irradiated equine herpesvirus type 1 (EHV-1). Biochemical transformants were isolated in a system selective for the dTK+ phenotype (Eagle minimal essential medium containing 10(-4) M hypoxanthine, 6 X 10(-7) M aminopterin, and 2 X 10(-5) M deoxythymidine). Transformation was accompanied by the acquisition of a dTK activity with immunological, electrophoretic, and biochemical characteristics identical to those of the dTK induced by EHV-1 during productive infection. The transformed cells have been maintained in selective culture medium for more than 50 passages and have retained the capacity to express EHV-1--specific antigens. Spontaneous release of infectious virus has not been detected in the transformed lines, and the the cells were not oncogenic for athymic nude mice. In contrast to normal dTk+ 3T3 cells, EHV-1 transformants were unable to grow in the presence of arabinosylthymine, a drug selectively phosphorylated by herpesvirus-coded dTK's. These results indicate that a portion of the EHV-1 genome is able to persist in the transformed cells for many generations and be expressed as an enzymatically active viral gene product.
缺乏脱氧胸苷激酶(dTK-)的3T3小鼠细胞系在暴露于紫外线照射的1型马疱疹病毒(EHV-1)后,稳定地转化为dTK+表型。在对dTK+表型具有选择性的系统(含有10^(-4)M次黄嘌呤、6×10^(-7)M氨基蝶呤和2×10^(-5)M脱氧胸苷的伊格尔基本培养基)中分离出生化转化体。转化伴随着获得一种dTK活性,其免疫学、电泳和生化特性与EHV-1在生产性感染期间诱导的dTK相同。转化细胞已在选择性培养基中传代培养50多次,并保留了表达EHV-1特异性抗原的能力。在转化细胞系中未检测到传染性病毒的自发释放,并且这些细胞对无胸腺裸鼠不具有致癌性。与正常的dTk+ 3T3细胞相反,EHV-1转化体在阿糖胸苷存在下无法生长,阿糖胸苷是一种由疱疹病毒编码的dTK选择性磷酸化的药物。这些结果表明,EHV-1基因组的一部分能够在转化细胞中持续存在许多代,并作为一种具有酶活性的病毒基因产物表达。