Transfection of mouse cells with thymidine kinase gene of herpes simplex virus

Chee Gun Lee, Chan Gil Kim, Rock Namkung, Sang Eun Lee, Sang Dai Park

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

A mouse cell line (LP1-1) was established from the murine L cells deficient in thymidine kinase (L-M(TK-)) by prolonged selective culture on the hypoxanthine-aminopterine-thymidine (HAT) medium following transfection with the thymidine kinase gene of herpes simplex virus type-I (HSV TK). Southern blot analysis has shown that the viral TK gene was integrated into one of the chromosomal loci by a single copy. From this established cell line, the 5-bromo-2-deoxyuridine (BrdU) resistant revertant was brought out at a frequency of 1×10-6 and from these BrdU resistant revertants (LP1BU), one out of 1×105 cells could return to the HAT-resistant phenotype. The established LP1-1 cell line showed a typical biphasic nature of DNA synthesis as determined by the 3H-thymidine incorporation test. The activity of thymidine kinase was shown to be equivalent to that of the DNA polymerase-α when the whole nuclear fraction or the nuclear matrix were used for examination. These results indicate that the transfected viral TK gene can be expressed under the normal cell-cycle regulation and its gene product can act as a component of the multienzyme complex which is responsible for DNA replication.

Original languageEnglish (US)
Pages (from-to)141-147
Number of pages7
JournalCytotechnology
Volume3
Issue number2
DOIs
StatePublished - Mar 1 1990
Externally publishedYes

Keywords

  • DNA polymerase-α
  • L-M(TK)
  • multienzyme complex
  • nuclear matrix
  • pHSV-106
  • thymidine kinase

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Clinical Biochemistry
  • Cell Biology

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