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Production of transgenic chickens expressing a tetracycline-inducible GFP gene
Authors:Kwon Mo Sun  Koo Bon Chul  Roh Ji Yeol  Kim Minjee  Kim Jin-Hoi  Kim Teoan
Institution:aDepartment of Physiology, Catholic University of Daegu School of Medicine, Daegu, Republic of Korea;bDepartment of Animal Sciences, Chungbuk National University, Cheongju, Republic of Korea;cDepartment of Animal Biotechnology, Konkuk University, Seoul, Republic of Korea
Abstract:There is much interest in using farm animals as ‘bioreactors’ to produce large quantities of biopharmaceuticals. However, uncontrolled constitutive expression of foreign genes have been known to cause serious physiological disturbances in transgenic animals. The objective of this study was to test the feasibility of the controllable expression of an exogenous gene in the chicken. A retrovirus vector was designed to express GFP (green fluorescent protein) and rtTA (reverse tetracycline-controlled transactivator) under the control of the tetracycline-inducible promoter and the PGK (phosphoglycerate kinase) promoter, respectively. G0 founder chickens were produced by infecting the blastoderm of freshly laid eggs with concentrated retrovirus vector. Feeding the chickens obtained with doxycycline, a tetracycline derivative, resulted in emission of green body color under fluorescent light, and no apparent significant physiological dysfunctions. Successful germline transmission of the exogenous gene was also confirmed. Expression of the GFP gene reverted to the pre-induction levels when doxycycline was removed from the diet. The results showed that a tetracycline-inducible expression system in transgenic animals might be a promising solution to minimize physiological disturbances caused by the transgene.
Keywords:Transgenic chicken  Green fluorescent protein (GFP)  Retrovirus vector  Tetracycline-inducible expression  Doxycycline
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