Kuo C H, Uetsuki T, Kim C H, Tanaka H, Li B S, Taira E, Higuchi H, Okamoto H, Yoshikawa K, Miki N
Department of Pharmacology I, Medical School, Osaka, Japan.
Biochem Biophys Res Commun. 1995 Jun 15;211(2):438-46. doi: 10.1006/bbrc.1995.1833.
To determine cis-acting elements required for neuron specific expression of a necdin gene, we tried to use zebra fish assay system in vivo instead of cell lines in vitro. Various expression vectors carrying upstream sequences of necdin gene fused to MEKA (lacZ) gene as a reporter were injected into fertilized zebra fish embryos and then the expression of the reporter gene was analyzed by the whole mount immunochemical method. No promoter activity was obtained with a construct carrying sequence from -63 to +63 of the necdin gene, while promoter activity with preferential skin expression was obtained with a construct having sequence from -86 to +28. Further upstream sequence from -173 to +28 exhibited neuron specific expression as well as that from -845 to +63. These results indicate that a cis-acting element responsible for neuron specific expression is located in an 87bp sequence from -173 to -87 of necdin gene.
为了确定神经细胞黏附分子（necdin）基因神经元特异性表达所需的顺式作用元件，我们尝试在体内使用斑马鱼检测系统而非体外细胞系。将各种携带与MEKA（lacZ）基因融合的necdin基因上游序列作为报告基因的表达载体注射到受精的斑马鱼胚胎中，然后通过整体免疫化学方法分析报告基因的表达。携带necdin基因从-63至+63序列的构建体未获得启动子活性，而携带从-86至+28序列的构建体获得了优先在皮肤表达的启动子活性。从-173至+28的更上游序列以及从-845至+63的序列均表现出神经元特异性表达。这些结果表明，负责神经元特异性表达的顺式作用元件位于necdin基因从-173至-87的87bp序列中。
