The regulation of GFP expression by Tet-on system in zebrafish heart
Date Issued
2004
Date
2004
Author(s)
Jeng, Yu-Ting
DOI
zh-TW
Abstract
The tetracycline (Tet) –controlled system is well developed in many transgenic organisms. Yet, this system has not been reported in zebrafish. DNA construct, rtTA that is driven by cmlc2 promoter acts as an activator of TetO, was microinjected into the one-celled stage zebrafish embryos and there of germ-lines were generated. We examined the germ-line transmitted F2 embryos; we found that the increase of GFP expression was tightly dependent on the higher Dox dosage (ranged from 10, 1, 0.1 to 0.01 ug/ml) and on the longer induction time (ranged from 12, 24, 36, 48, 60, 72, 84 to 96 hr). When the 18-hpf embryos that de-chorion and the 24-hpf embryos were treated with 10 ug/ml Dox, GFP was specifically expressed in heart after 7 hr induction. When the 10-week fish were immersed in 10 ug/ml Dox, then GFP expression were expressed specifically in the heart after 24 hr induction. Meanwhile, the 3-dpf embryos were treated with 10 ug/ml Dox for 1 hr, the mean intensity of GFP increased gradually after withdrawal of Dox for 3 days. But after withdrawal of Dox for 5 days GFP signal disappeared completely. Therefore, Tet-ON system is capable of being turned down if Dox was removed. These evidences are the first report to demonstrate that the tet-on system can work in transgenic model fish and is highly potentially useful to study gene function in specific tissue at any stages whenever gene is induced.
Subjects
斑馬魚
誘導式調控
Doxycycline
zebrafish
Tet-on system
Type
other
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