8‐Br‐cAMP inhibits the transient expression of firefly luciferase

FEBS Letters

Coker, G.T.¹  O'Malley, K.L.¹  Vinnedge, L.¹ 

[1] Department of Anatomy and Neurobiology, Washington University Medical School, St. Louis, MO 63110, USA

关键词: Luciferase;    Transfection;    Bromo cyclic AMP;    8-;    Reporter gene;    Methyl-3-isobutylmethylxanthine;    1-;    CAT;    chloramphenicol acetyltransferase;    IBMX;    1-methyl-3-isobutylxanthine;    TH;    tyrosine hydroxylase;   

DOI  :  10.1016/0014-5793(89)80620-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The genes for firefly luciferase and chloramphenicol acetyltransferase (CAT) were used as reporter genes to explore the activation of heterologous promoters by 8-Br-cAMP. Cells were transfected with a CAT gene/tyrosine hydroxylase promoter, which contains a cAMP response element. Extracts from cells treated with 8-Br-cAMP had 340% more enzyme activity than untreated cells. In contrast, treated cells transfected with a tyrosine hydroxylase/luciferase construct had 30% less activity than control cells. Simian virus and rous sarcoma virus promoters/luciferase constructs also had lower activities in cells treated with 8-Br-cAMP than untreated cells. The inhibition of luciferase enzyme activity by cAMP appears to be posttranscriptional since both luciferase and CAT RNA levels were similarly increased in cells treated with 8-Br-cAMP or 1-methyl-3-isobutylmethylxanthine. The lower level of luciferase activity was not due to simple allosteric inhibition. We conclude that constructs using the firefly luciferase as a reporter gene are unsuitable for studying the effects of cAMP on the regulation of promoters.

【 授权许可】

Unknown