Construction and characterization of a recombinant plasmid encoding the gene for the thymidine kinase of herpes simplex type 1 virus

Lynn W. Enquist, George F. Vande Woude, Michael Wagner, James R. Smiley, William C. Summers

Research output: Contribution to journalArticlepeer-review

79 Scopus citations

Abstract

We have constructed a hybrid plasmid by insertion of the thymidine kinase (TK) gene of Herpes simplex virus (HSV) type I at the BamHI site on Escherichia coli plasmid pBR322. The restriction endonuclease cleavage site map for the viral DNA fragment was determined for ten nucleases, and the insert in the recombinant plasmid has the same restriction nuclease digestion pattern as bona fide viral DNA. This result indicates that the plasmid contains an accurate copy of the viral DNA. The viral TK gene carried on the plasmid can be introduced into mammalian cells where it is expressed. This source of DNA with a selectable marker should be of considerable practical use in gene-transfer experiments in mammalian cells.

Original languageEnglish (US)
Pages (from-to)335-342
Number of pages8
JournalGene
Volume7
Issue number3-4
DOIs
StatePublished - Nov 1979

All Science Journal Classification (ASJC) codes

  • Genetics

Keywords

  • Recombinant DNA
  • biochemical transformation
  • cloning
  • herpes virus
  • restriction endonuclease site maps

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