Expression of herpesvirus thymidine kinase gene under control of early promoter of SV40

A 0.31-kilobase fragment of SV40 DNA containing the early promoter of this virus was isolated. The pX1 plasmid containing the herpesvirus thymidine kinase gene (TK) was cleaved with BglII and SalI to remove the TK promoter. The Sv40 DNA fragment was mixed with the TK gene without its promoter and blunt-end ligated after treatment with S₁ nuclease. The newly engineered plasmid was used for the transformation of TK-deficient LtA cells to TK⁺ phenotype. Blot hybridization experiments show that the plasmid is integrated into high-molecular-weight DNA in both of the transformed colonies that we have studied. Also the mRNA in these cells is a hybrid molecule containing both TK and SV40 sequences.