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외래 유전자의 효율적인 발현 조절을 위한 개선된 Tetracycline-Inducible Expression System의 구축
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  • 외래 유전자의 효율적인 발현 조절을 위한 개선된 Tetracycline-Inducible Expression System의 구축
  • Construction of Improved Tetracycline-Inducible Expression System for the Effective Regulation of Transgene Expression
저자명
구본철,권모선,김태완,Koo. Bon-Chul,Kwon. Mo-Sun,Kim. Teo-An
간행물명
Reproductive & developmental biology
권/호정보
2009년|33권 1호|pp.63-69 (7 pages)
발행정보
한국동물번식학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

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.