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Regulation of GFP Expression Using the Tetracycline Inducible Retroviral Vector System  

Koo Bon Chul (Department of Physiology, Catholic University of Daegu School of Medicine)
Kwon Mo Sun (Department of Physiology, Catholic University of Daegu School of Medicine)
Kim Teoan (Department of Physiology, Catholic University of Daegu School of Medicine)
Publication Information
Reproductive and Developmental Biology / v.29, no.1, 2005 , pp. 57-62 More about this Journal
Abstract
One of the critical problems to be solved in transgenic animal production is uncontrollable constitutive expression of foreign genes, which usually results in serious physiological disturbances in the transgenic animal. To circumvent this problem, we constructed and tested two retrovirus vectors designed to express the GFP(green fluorescent protein) gene under the control of the tetracycline-inducible promoters. To maximize the GFP gene expression at turn-on state, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was introduced into the retrovirus vectors at downstream region of either the GFP gene or the sequence encoding rtTA(reverse tetracycline-controlled transactivator). Transformed cells were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the GFP gene expression level using fluorometry and western blotting. Higher GFP expression was observed from the vector carrying the WPRE sequence at 3' side of the GFP gene, while tighter expression control(up to 20 fold) was obtained from the vector in which the WPRE sequence was placed at 3' side of rtTA sequence. The resulting tetracycline inducible vector system may be used in transgenic animal production and gene therapy.
Keywords
rtTA system; WPRE; GFP; Doxycycline; Induction efficiency;
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