Schilling K, Luk D, Morgan J I, Curran T
Department of Neurosciences, Roche Institute of Molecular Biology, Roche Research Center, Nutley, NJ 07110.
Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5665-9. doi: 10.1073/pnas.88.13.5665.
Expression of the c-fos protooncogene is induced by a great variety of extracellular stimuli. A fos-lacZ fusion gene has been constructed that recapitulates this regulation. The fos-lacZ gene was introduced into B104 neuroblastoma cells for use in a quantitative assay for stimulus-transcription coupling. Both alpha- and beta-adrenergic agonists, dibutyryl cAMP, and phorbol ester induced beta-galactosidase activity in a dose-dependent manner. Thus, the interactions of receptors with agonists and antagonists, as well as intracellular second messenger-mediated signaling events, can be analyzed quantitatively. This approach represents a prototypic method for investigating stimulus-response coupling based upon gene expression.
原癌基因c-fos的表达可由多种细胞外刺激诱导产生。现已构建了一个能重现这种调控作用的fos-lacZ融合基因。将fos-lacZ基因导入B104神经母细胞瘤细胞，用于刺激-转录偶联的定量检测。α-和β-肾上腺素能激动剂、二丁酰环磷腺苷及佛波酯均可剂量依赖性地诱导β-半乳糖苷酶活性。因此，可对受体与激动剂及拮抗剂的相互作用，以及细胞内第二信使介导的信号转导事件进行定量分析。这种方法代表了一种基于基因表达来研究刺激-反应偶联的原型方法。
