McGowan J J, Allen G P, Gentry G A
Infect Immun. 1979 Aug;25(2):610-4. doi: 10.1128/iai.25.2.610-614.1979.
A line of mouse 3T3 cells lacking deoxythymidine kinase (dTK-) was stably transformed to see dTK+ phenotype after exposure to ultraviolet-irradiated equine herpesvirus type 1 (EHV-1). Deoxythymidine kinase (dTK) was purified from the biochemically transformed mouse cells by affinity chromatography on deoxythymidine-Sepharose. The purified dTK from EHV-1-transformed 3T3 cells was identical to the dTK purified from dTK- 3T3 cells lytically infected with EHV-1 with respect to its electrophoretic mobility, molecular weight, substrate specificity, phosphate donor specificity, and immunological specificity. The sedimentation velocity of the purified dTK from the transformed 3T3 cells was similar to that previously reported for the enzyme in lytically infected dTK- 3T3 cells, and its molecular weight was estimated to be 87,000. Antiserum prepared against the EHV-1 dTK induced in horse cells inactivated the dTK purified from the transformed mouse cells. The Km for deoxythymidine (5 micrometers) of purified dTK from the EHV-1-transformed cells was the same as that reported for the EHV-1-induced dTK. These results further support the notion that the dTK acquired by dTK- mouse 3T3 cells after transformation by EHV-1 is of viral and not of cellular origin.
将缺乏脱氧胸苷激酶（dTK-）的小鼠3T3细胞系进行稳定转化，使其在暴露于紫外线照射的1型马疱疹病毒（EHV-1）后呈现dTK+表型。通过在脱氧胸苷-琼脂糖上进行亲和层析，从经生化转化的小鼠细胞中纯化出脱氧胸苷激酶（dTK）。从EHV-1转化的3T3细胞中纯化得到的dTK，在电泳迁移率、分子量、底物特异性、磷酸供体特异性和免疫特异性方面，与从被EHV-1裂解感染的dTK- 3T3细胞中纯化得到的dTK相同。从转化的3T3细胞中纯化得到的dTK的沉降速度，与先前报道的在被裂解感染的dTK- 3T3细胞中的该酶的沉降速度相似，其分子量估计为87,000。针对在马细胞中诱导产生的EHV-1 dTK制备的抗血清，使从转化的小鼠细胞中纯化得到的dTK失活。从EHV-1转化的细胞中纯化得到的dTK对脱氧胸苷的Km（5微摩尔），与报道的EHV-1诱导的dTK的Km相同。这些结果进一步支持了这样一种观点，即dTK-小鼠3T3细胞在被EHV-1转化后获得的dTK是病毒来源而非细胞来源。
