High efficiency transient expression of eukaryotic genes: use of an HSV-1 immediate early promoter (ICP4).
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 chloramphenicol 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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West Virginia University Health Sciences Center.
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D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D017874:Immediate-Early Proteins; D008114:Liver Neoplasms, Experimental; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051381:Rats; D018139:Simplexvirus / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics*
D017874:Immediate-Early Proteins; D010750:Phosphoproteins; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C048925:herpes simplex virus, type 1 protein ICP4; D015500:Chloramphenicol O-Acetyltransferase