New doxycycline-inducible transgenic lines in Xenopus.

New doxycycline-inducible transgenic lines in Xenopus.
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DOI:
10.1002/dvdy.22642
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发表时间:
2011-06
影响因子:
2.5
通讯作者:
Buchholz, Daniel R.
Buchholz, Daniel R.
中科院分区:
生物学3区
文献类型:
--
作者:
Rankin, Scott A.;Zorn, Aaron M.;Buchholz, Daniel R.

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我们利用Tet-on可诱导系统鉴定了两个新的能够实现转基因表达的转基因非洲爪哇品系。诱导系在无处不在的启动子CMV的控制下表达多西环素(Dox)激活的转录因子RTTA。响应系能够通过四环素响应性转基因启动子(TrE)诱导Dox诱导的显性阳性甲状腺激素受体的表达。DOX诱导的转基因GFP基因表达在3小时后即可检测到,2天后可增加10~50倍,这取决于Dox的剂量。在Dox去除后3d内,诱导的GFP mRNA表达恢复到未诱导的水平。用Dox处理RTTA诱导子和TrE响应子双转基因动物,可加速甲状腺激素反应基因表达和形态的变性改变。这些转基因品系将通过新的非洲爪哇种群中心提供,将成为发育和变态遗传分析的宝贵工具。
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 hours 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.
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