Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Construction of Improved Tetracycline-Inducible Expression System for the Effective Regulation of Transgene Expression

외래 유전자의 효율적인 발현 조절을 위한 개선된 Tetracycline-Inducible Expression System의 구축

  • 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, Teo-An (Department of Physiology, Catholic University of Daegu School of Medicine)
  • 구본철 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 권모선 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 김태완 (대구가톨릭대학교 의과대학 생리학교실)
  • Published : 2009.03.31
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Abstract

In this study we tried to construct a more efficient tetracycline-inducible gene expression system by replacing previous key elements with more advance ones. At the beginning, we substituted PGK (phophoglycerate kinase) promoter for CMV (cytomegalovirus) promoter to control "$rtTA2^sM2$" which has been known for high induction efficiency in response to tetracycline. With this modification, expression of the EGFP marker gene under the induction condition was significantly increased. Next, we replaced "TRE" fragment with a modified version named "TRE-tighf" which has been reported to have higher affinity and specificity to the transactivator by minor base change of the "TRE" DNA fragment sequence. Use of "TRE-tighf" instead of "TRE" resulted in more than 10 fold increment in terms of induction efficiency and significant decrement of background expression in non-inducible condition. By combining PGK promoter and "TRE-tight" fragment, we could upgrade previous tetracycline-inducible system to show more stringent turn on/off gene switch ability and stronger expression of the gene of our interest. Use of this newly developed system must be very helpful to the studies of gene expression, especially to the transgenic animal study in which non-controllable constitutive expression of the transgene has been one of the urgent problems to be solved.

Keywords

References

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