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http://dx.doi.org/10.12749/RDB.2013.37.3.109

Examination of Improved Tetracycline Inducible Gene Expression System In Vitro  

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)
Koo, Bon Chul (Department of Physiology, Catholic University of Daegu School of Medicine)
Publication Information
Reproductive and Developmental Biology / v.37, no.3, 2013 , pp. 109-115 More about this Journal
Abstract
Until recently the most popular tetracycline-inducible gene expression system has been the one developed by Gossen and Bujard. In this study, we tested the latest version of same system and the results are summarized as follows: Compared with previous one, the difference of new system are minor changes of nucleotide sequences in transactivator and tetracycline response element (TRE) regions. Sensitivity to the doxycycline (a tetracycline derivative) was improved. Leakiness of GFP marker gene expression in non-inducible condition was significantly decreased. Higher expression of the marker gene was observed when the cells were fed with doxycycline-containing medium. Optimal insertion site of woodchuck posttranscriptional regulatory element (WPRE) sequence which was known to increase gene expression was different depending on the origin of cells. In chicken embryonic fibroblast, location of WPRE sequence at 3' end of TRE resulted in the highest GFP expression. In bovine embryonic fibroblasts, 3' end of transactivator was the best site for the GFP expression.
Keywords
Tet system; Transactivator; TRE; WPRE; Primary culture cell;
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