Characterization of a Novel Type I Keratin Gene and Generation of Transgenic Lines With Fluorescent Reporter Genes Driven by Its Promoter/Enhancer in Xenopus laevis
Characterization of a Novel Type I Keratin Gene and Generation of Transgenic Lines With Fluorescent Reporter Genes Driven by Its Promoter/Enhancer in Xenopus laevis
复制标题
DOI：
10.1002/dvdy.22451
复制
发表时间：
2010-12-01
影响因子：
2.5
通讯作者：
Mochii, Makoto
中科院分区：
文献类型：
--
作者：
Suzuki, Ken-Ichi T.;Kashiwagi, Keiko;Mochii, Makoto
我们研究了非洲爪蟾个体发生过程中一个新的I型角蛋白基因的特征。在神经发育后期，转录本首先在鱼的后部被检测到，然后在胚胎发育期间局限于鳍和外鳃。为了研究角蛋白基因在体内的转录调控作用，我们构建了由其4.2 kb上游序列驱动的荧光报告基因转基因系。启动子/增强子活性重现了胚胎发生过程中内源基因的表达。序列缺失分析显示，启动子附近的区域对鱼鳍特异性表达至关重要。报告基因在包括鳍和鳃在内的多个器官中均有表达。特别是在发育中的肢体和鳃中观察到强烈的表达。在变态过程中，报告荧光随着鳃内吸收迅速下降。携带启动子/增强子的转基因品系将为阐明各种器官，特别是鳃的形成、发育和吸收提供有价值的工具。生态学报，2009(3):379 - 381。(C) 2010 Wiley-Liss, Inc。
We investigated the characteristics of a novel type I keratin gene in Xenopus laevis during ontogenesis. The transcript was first detected in the posterior region at the late neurula stage, and then restricted to the fin and external gill during embryogenesis. To examine the transcriptional regulation of the keratin gene in vivo, we generated transgenic lines with fluorescent reporter genes driven by its 4.2-kb upstream sequence. The promoter/enhancer activity recapitulated the endogenous gene expression during embryogenesis. Sequential deletion analyses revealed that the regions proximal to the promoter were essential for fin-specific expression. Reporter expression was detected in various organs, including the fin and gill. In particular, robust expression was observed in the developing limbs and gill. The reporter fluorescence rapidly decreased with internal gill resorption during metamorphosis. The transgenic lines carrying the promoter/enhancer should represent valuable tools for elucidating the formation, development and resorption of various organs, especially the gill. Developmental Dynamics 239:3172-3181, 2010. (C) 2010 Wiley-Liss, Inc.