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Nucleic Acids Research, 1979, Vol. 7, No. 4 859-878
© 1979

Articles

Hybrid plasmids containing an active thymidine kinase gene of Herpes simplex virus 1

N.M. Wilkie{dagger},+, J.B. Clements+, W. Boll*, N. Mantei*, D. Lonsdale+ and C. Weissmann*

+Institute of Virology, University of Glasgow Church Street, Glasgow G11 5JR, Scotland *Institut für Molekularbiologie I, Universität Zürich 8093 Zürich, Switzerland

Received August 7, 1979.

The gene for the thymidine kinase (TK) of Herpes simplex virus type 1 (HSV-1) is located in the KpnI m and BamHI P fragments of the genome (Wigler et al., Cell 11, 223–232 (1977)). These fragments have been inserted into the EcoRI and BamHI sites, respectively, of plasmid pBR322 propagated in E.coli. The TK gene contained in the recombinant plasmids was shown to be biologically active when introduced into TK– mouse L cells. Detailed restriction site maps of the BamHI p fragment have been constructed and the approximate location of the TK gene has been determined. Mouse cells transformed with cloned HSV-1 tk+ DNA produced HSV-1-specific thymidine kinase; superinfection with HSV-1 tk– virus increased the level of TK activity tenfold, suggesting that the BamHI p sequences present in transformed cells respond to virus-encoded regulatory gene product(s).

{dagger}N.M.Wilkie is a member of the Medical Research Council Virology Unit.


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