Discordant estimates of heterologous promoter activity as determined by reporter gene mRNA levels and enzyme activity.
Biochem Biophys Res Commun
; 189(2): 912-8, 1992 Dec 15.
Article
em En
| MEDLINE
| ID: mdl-1335248
ABSTRACT
In this study, the human cytomegalovirus (CMV) promoter fused to the lacZ (beta-gal) reporter gene was transfected into neuroblastoma SK-N-BE(2)-C cells, and phorbol ester-stimulated promoter activity assessed by both PCR quantitation of reporter gene mRNA levels and enzyme activity. Surprisingly, significant differences were observed in the induction profile of CMV promoter activity as judged by these two independent methods of analysis. For example, at 24 hrs post-transfection beta-gal activity was elevated 7.3-fold in phorbol ester-treated cells, whereas 2.4-fold increases were observed in the cognate mRNA levels. These findings demonstrate the efficacy of quantitative PCR methodology to evaluate promoter activity in DNA-mediated cell transfection analyses, and raise a cautionary note on the reliance of reporter gene enzyme activity to estimate the transcriptional activity of heterologous promoters.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes
/
RNA Mensageiro
/
Cloranfenicol O-Acetiltransferase
/
Regiões Promotoras Genéticas
/
Beta-Galactosidase
/
Citomegalovirus
Limite:
Humans
Idioma:
En
Revista:
Biochem Biophys Res Commun
Ano de publicação:
1992
Tipo de documento:
Article