Biochemical transformation of mouse cells by fragments of herpes simplex virus DNA

Abstract

Mouse L cells lacking the enzyme thymidine kinase (LMTK-) have been converted to a TK+ phenotype by infection with fragmented HSV2 strain 333 DNA. The DNA fragments used were either unique, produced by cleavage with the restriction endonucleases Eco RI and Hind III, or randomly produced by mechanical shearing. Survival in HAT medium was used initially to establish the TK+ phenotype; clones possessing the ability to grow in selective medium were picked on the basis of differing morphology and growth rates. Cytosol extracts of these clones possessed virus-specified TK activity identical to that present in cells lytically infected with HSV2, as indicated by thermolability and mobility on polyacrylamide gel electrophoresis. The transformed cells also exhibit HSV-specific immunofluorescence. Based on these transformation studies, it is possible to assign a map location to the TK gene on the HSV genome.