High efficiency transient expression of eukaryotic genes: use of an HSV-1 immediate early promoter (ICP4).
Biotechniques
; 9(2): 168-73, 1990 Aug.
Article
en En
| MEDLINE
| ID: mdl-2169263
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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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Fosfoproteínas
/
Transfección
/
Transactivadores
/
Regulación de la Expresión Génica
/
Regiones Promotoras Genéticas
/
Simplexvirus
/
Proteínas Inmediatas-Precoces
/
Proteínas Reguladoras y Accesorias Virales
Límite:
Animals
Idioma:
En
Revista:
Biotechniques
Año:
1990
Tipo del documento:
Article
Pais de publicación:
Reino Unido