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

5 Citations (Scopus)

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

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Human Herpesvirus 1
Viruses
Genes
Gene Expression
Reporter Genes
Transfection
Herpes Simplex Virus Protein Vmw65
Plasmids
Bearings (structural)
Polyadenylation
Acetyltransferases
Cloning
Virion
Transcriptional Activation
Rats
Organism Cloning
Hepatocellular Carcinoma
DNA

All Science Journal Classification (ASJC) codes

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

Cite this

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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.",
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High efficiency transient expression of eukaryotic genes : Use of an HSV-1 immediate early promoter (ICP4). / Liu, X.; Giza, C. C.; Vrana, Kent.

In: BioTechniques, Vol. 9, No. 2, 27.08.1990, p. 168-173.

Research output: Contribution to journalArticle

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