Division of Developmental Biology, Cincinnati Children's Research Foundation and Department of Pediatrics, College of Medicine, University of Cincinnati, Cincinnati, Ohio 45208, USA.
Dev Dyn. 2011 Jun;240(6):1467-74. doi: 10.1002/dvdy.22642. Epub 2011 Apr 12.
We have characterized two new transgenic Xenopus lines enabling transgene expression using the Tet-On inducible system. An inducer line expresses the doxycycline- (Dox-) activated transcription factor rtTA under control of the ubiquitous promoter CMV. A responder line enables Dox-inducible expression of a dominant positive thyroid hormone receptor via a tetracycline responsive transgenic promoter (TRE). Dox-induced expression of transgenic GFP mRNA was detectable after 3 hr and increased up to 10- to 50-fold by 2 days depending on dose of Dox. Induced GFP mRNA expression returned to uninduced levels within 3 days upon Dox removal. Treatment of rtTA inducer and TRE responder double transgenic animals with Dox caused acceleration of metamorphic changes in thyroid hormone-response gene expression and morphology. These transgenic lines will be made available through the new Xenopus Stock Center and will serve as valuable tools for genetic analysis of development and metamorphosis.
我们已经鉴定了两个新的转基因爪蟾系，它们可以使用 Tet-On 诱导系统进行转基因表达。一个诱导系在 CMV 普遍启动子的控制下表达强力霉素（Dox）激活的转录因子 rtTA。一个应答系通过四环素反应性转基因启动子（TRE）使显性阳性甲状腺激素受体实现 Dox 诱导表达。在 3 小时后可以检测到 Dox 诱导的转基因 GFP mRNA 的表达，并根据 Dox 的剂量增加到 2 天的 10-至 50 倍。在 Dox 去除后 3 天内，诱导的 GFP mRNA 表达恢复到未诱导水平。用 Dox 处理 rtTA 诱导系和 TRE 应答系双转基因动物导致甲状腺激素反应基因表达和形态的变态变化加速。这些转基因系将通过新的爪蟾品系中心提供，并将成为发育和变态遗传分析的有价值的工具。
