FEBS Letters | |
8‐Br‐cAMP inhibits the transient expression of firefly luciferase | |
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
【 预 览 】
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