Tet-OneTM Inducible Expression System

Description

The Tet-One Inducible Expression System is a tetracycline-inducible gene expression system that allows you to establish a tightly inducible expression system for your gene of interest in mammalian cells using an all-in-one plasmid vector. The pTetOne Vector expresses the Tet-On 3G transactivator from the constitutive human PGK promoter in the forward orientation, and your gene of interest from thePTRE3GSpromoter in the reverse orientation. In the presence of doxycycline Dox , the Tet-On 3G transactivator specifically binds and activates high-level transcription from the inducible promoter that controls expression of your gene. In addition, the kit contains linear selection markers for resistance to puromycin and hygromycin, either of which can be cotransfected with the pTetOne Vector to obtain stable clones. It also includes Xfect Transfection Reagent and Tet System Approved FBS. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Back Tet-One System Mechanism The Tet-On 3G transactivator protein is expressed constitutively from the humanPGKpromoter PPGK but is unable to bind to theTRE3Gpromoter PTRE3G in the absence of doxycycline Dox .When bound by Dox, supplied in the culture medium, the transactivator undergoes a conformational change, binds toPTRE3Gand activates transcription of a transgene cloned downstream. Back Required Products Fetal Bovine Serum FBS that has been functionally tested for optimal use with all of our Tet Systems. This FBS does not contain trace levels of tetracycline or derivatives which have been observed to interfere with proper regulation of TRE-controlled gene expression. Use of this serum ensures the maximal range of induction possible with the Tet Systems. This FBS has been processed in the United States. Back Fetal bovine serum FBS approved for use with tetracycline inducible expression systems.Only Takara Bio USA functionally tests FBS to be sure that no contaminating tetracycline derivatives interfere with induced expression levels. Back Fetal bovine serum from Clontech is functionally tested to guarantee consistent results with tetracycline-inducible expression systems.Serum from other vendors that have not been functionally tested may contain tetracyclines that affect the maximum expression in Tet-Off systems, or generate background in Tet-On systems. Data here shows induction of luciferase expression from our CHO-AA8-Luc Tet-Off Control Cell Line using different sources of FBS. Back Fetal Bovine Serum FBS that has been functionally tested for optimal use with all of our Tet Systems. This FBS does not contain trace levels of tetracycline or its derivatives which have been observed to interfere with proper regulation of TRE-controlled gene expression. Use of this serum ensures the maximal range of induction possible with the Tet Systems. Back Fetal bovine serum FBS approved for use with tetracycline inducible expression systems.Only Takara Bio USA functionally tests FBS to be sure that no contaminating tetracycline derivatives interfere with induced expression levels. Back Fetal bovine serum from Clontech is functionally tested to guarantee consistent results with tetracycline-inducible expression systems.Serum from other vendors that have not been functionally tested may contain tetracyclines that affect the maximum expression in Tet-Off systems, or generate background in Tet-On systems. Data here shows induction of luciferase expression from our CHO-AA8-Luc Tet-Off Control Cell Line using different sources of FBS. Back