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一种新型 I 型角蛋白基因的鉴定及其启动子/增强子驱动荧光报告基因在非洲爪蟾中的转基因系的构建。

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.

机构信息

Department of Biological Science, Graduate School of Science, Hiroshima University, Hiroshima, Japan.

出版信息

Dev Dyn. 2010 Dec;239(12):3172-81. doi: 10.1002/dvdy.22451. Epub 2010 Oct 12.

Abstract

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.

摘要

我们研究了非洲爪蟾在个体发生过程中新型 I 型角蛋白基因的特征。该转录本首先在晚期神经胚期的后区被检测到,然后在胚胎发生过程中局限于鳍和外部鳃。为了在体内检查角蛋白基因的转录调控,我们使用由其 4.2kb 上游序列驱动的荧光报告基因生成了转基因系。启动子/增强子活性在胚胎发生过程中重演了内源性基因表达。连续缺失分析表明,靠近启动子的区域对于鳍特异性表达是必需的。在各种器官中检测到报告基因表达,包括鳍和鳃。特别是在发育中的四肢和鳃中观察到强烈的表达。在变态过程中随着内鳃吸收,报告荧光迅速降低。携带启动子/增强子的转基因系应该代表阐明各种器官,特别是鳃的形成、发育和吸收的有价值的工具。

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