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A functional analysis of ACP-20, an adult-specific cuticular protein gene from the beetle Tenebrio: role of an intronic sequence in transcriptional activation during the late metamorphic period

机译:甲虫黄粉虫的成年特异性表皮蛋白基因ACP-20的功能分析:内含子序列在变态后期的转录激活中的作用

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摘要

A gene encoding the adult cuticular protein ACP-20 was isolated in Tenebrio. It consists of three exons interspersed by two introns, intron 1 interrupting the signal peptide. To understand the regulatory mechanisms of ACP-20 expression, ACP-20 promoter-luciferase reporter gene constructs were transfected into cultured pharate adult wing epidermis. Transfection assays needed the presence of 20-hydroxyecdysone, confirming that ACP-20 is up-regulated by ecdysteroids. Analysis of 5' deletion constructs revealed that three regions are necessary for high levels of transcription. Interaction experiments between intronic fragments and epidermal nuclear proteins confirmed the importance of intron 1 in ACP-20 transcriptional control, which results from the combined activity of regulatory cis-acting elements of the promoter and those of intron 1. [References: 94]
机译:在黄粉虫中分离出编码成年表皮蛋白ACP-20的基因。它由三个外显子组成,两个外显子散布着两个内含子,内含子1中断了信号肽。为了理解ACP-20表达的调节机制,将ACP-20启动子-荧光素酶报告基因构建体转染到培养的pha状成年翼表皮中。转染测定需要20-羟基蜕皮激素的存在,从而确认ACP-20被蜕皮类固醇上调。 5'缺失构建体的分析表明,三个区域对于高水平的转录是必需的。内含子片段与表皮核蛋白之间的相互作用实验证实了内含子1在ACP-20转录控制中的重要性,这是由于启动子和内含子1的顺式作用元件的结合活性共同引起的。[参考文献:94]

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