Dinces N B, Milman G
J Virol. 1982 May;42(2):691-9. doi: 10.1128/JVI.42.2.691-699.1982.
A recombinant DNA of 5,150 base pairs was prepared containing the intact early region of polyoma virus, including the viral origin of replication and the structural sequences of the herpes simplex virus type 1 thymidine kinase gene. Although no thymidine kinase activity was detected when herpes structural sequences alone were transfected into cells, activity was produced when the structural gene followed the polyoma early region. The recombinant DNA was encapsidated into polyoma virions when cotransfected into mouse 3T6 cells with helper DNA from an early polyoma virus mutant. Herpes thymidine kinase activity was detected by a rapid in situ autoradiographic assay in which [125]iododeoxycytidine was utilized as a substrate for the viral but not the cellular enzyme.
制备了一个5150个碱基对的重组DNA，其包含多瘤病毒完整的早期区域，包括病毒复制起点和单纯疱疹病毒1型胸苷激酶基因的结构序列。虽然单独将疱疹结构序列转染到细胞中时未检测到胸苷激酶活性，但当结构基因位于多瘤病毒早期区域之后时则产生了活性。当与来自早期多瘤病毒突变体的辅助DNA共转染到小鼠3T6细胞中时，重组DNA被包装到多瘤病毒颗粒中。通过一种快速原位放射自显影测定法检测疱疹胸苷激酶活性，其中使用[125]碘脱氧胞苷作为病毒而非细胞酶的底物。
