High efficiency transient expression of eukaryotic genes: Use of an HSV-1 immediate early promoter (ICP4)

X. Liu, C. C. Giza, Kent Vrana

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

We have compared the transcriptional efficiencies of a number of eukaryotic promoters following DNA-mediated transfection into cultured rat hepatoma cells. We find that the highest levels of expression for the bacterial chloramphenical acetyltransferase (CAT) reporter gene are observed with a herpes simplex virus type 1 (HSV-1) immediate early promoter when co-transfected with an expression construct bearing the gene for the HSV-1 transcriptional activator protein VP16. This transactivation phenomenon is specific for the HSV-1 immediate early promoter and increases the expression of the reporter gene 7-fold. Expression from the ICP4 promoter is 2.5-fold greater than the other promoters tested. In addition, expression from the ICP4 promoter can be induced, at varying times following transfection, by infecting the cells with HSV-1 viral particles. Two plasmids have been constructed which contain the HSV-1 ICP4 promoter adjacent to a multiple cloning site. One of the plasmids also contains SV40 splicing and polyadenylation signals.

Original languageEnglish (US)
Pages (from-to)168-173
Number of pages6
JournalBioTechniques
Volume9
Issue number2
StatePublished - Aug 27 1990

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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