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.

ABSTRACT   
서론   
실험 방법    
새로운 Tet System의 구축    
Virus의 생산 및 표적세포로의 감염    
형관현미경 하에서 
Tet System에 의한 EGFP 유전자 발현관찰    
RT-PCR 분석    
Western Blotting 분석    
ELISA 분석   
결과    
형광현미경 하에서의 각 Vector에 따른 EGFP 유전자의 발현 조절 양상 관찰    
각 세포주에서 EGFP 유전자의 전이에 대한 RT-PCR 분석    
각 세포주에서 EGFP 발현에 대한 단백질 수준의 정량 분석    
각 Tet System의 Doxycycline에 대한 민감도 확인   
고찰   
인용문헌

• 권모선(대구가톨릭대학교 의과대학 생리학교실) | Mo Sun Kwon
• 김태완(대구가톨릭대학교 의과대학 생리학교실) | Teoan Kim
• 구본철(대구가톨릭대학교 의과대학 생리학교실) | Bon Chul Koo Corresponding author