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Inducible System · Yeast hybrid
Tetracycline 유도 발현 시스템
> Tet-Express – Fast, Simple Tet-Inducible Expression
Tet-Express – Fast, Simple Tet-Inducible Expression
Tet-Express - Fast, Simple Tet-Inducible Expression
Fast to set up
- no need to create a Tet-On or Tet-Off cell line
Fast to induce
- reach 80% of maximal expression in just 2 hours
- instead just add Tet-Express transactivator directly to cells
Clontech's Tet-Express Inducible Expression Systems are a faster, simpler adaptation of our powerful, tightly-regulated Tet-On/Tet-Off expression systems (1-3). Unlike Tet-On/Tet-Off, the Tet-Express system requires only a single vector, the Tet-Express transducible protein, and a doxycycline-free protocol. To induce expression, simply apply a few microliters of Tet-Express to the culture medium of cells in which your gene is under the control of any TRE-containing promoter. Tet-Express makes tetracycline-controlled transcription a rapid process for all cell types, and is particularly advantageous for cell types that are not amenable to sequential rounds of clonal selection.
What is Tet-Express?
Tet-Express is a version of Clontech’s Tet-Off Advanced transactivator protein that has been modified and optimized for self-transduction, i.e., it has the ability to transport itself across cell membranes into the nucleus via protein transduction pathways (Figure 1). Since Tet-Express binds and activates expression in the absence of tetracyclines, doxycycline is not required for gene activation. An Intensifier Reagent included with each vial of Tet-Express increases protein transduction efficiency.
The Tet-Express System, like Clontech’s Tet-On 3G System, expresses your transgene from a vector containing the tightly-regulated inducible promoter PTRE3G. However, unlike the Tet-On 3G system, you do not need to create a double-stable cell line that expresses the tetracycline transactivator, since the self-transducing Tet-Express transactivator protein is added directly to your cells.
Low Basal Expression, High Maximal Expression & Rapid Induction
A pTRE3G vector is included with every Tet-On 3G and Tet-Express system. This vector contains the inducible promoter (P
), the most advanced TRE-containing promoter available, which has been mutated to ensure very low basal expression and high maximal expression after induction (4; see www.clontech.com/teton3g). Because PTRE3G lacks binding sites for endogenous mammalian transcription factors, and since the minimal CMV promoter is inactive in the absence of a tetracycline transactivator protein, basal expression from P
is the lowest of any TRE-containing promoter available so far. Basal expression is undetectable by Western analysis, but expression can be induced to very high levels in the presence of a tetracycline transactivator (Figure 2). The Tet-Express System provides rapid induction of high levels of gene expression in a wide variety of cell types, as described in our Tet-Express technical note.
Choice of Four Different Tet-Express System Formats
The Tet-Express Inducible Expression System (which contains the core pTRE3G vector) is also available in three other vector formats (Figure 3). These include the Tet-Express Inducible Expression System (Bicistronic Version), which allows inducible coexpression of two genes of interest; and the Tet-Express Inducible Expression System (with mCherry) and the Tet-Express Inducible Expression System (with ZsGreen1), which allow monitoring of inducibility via induced coexpression of a red (mCherry) or green (ZsGreen1) fluorescent protein. Each format is sold as a complete system (see the Components section below for more information).
Figure 2. Basal expression from the PTRE3G promoter is undetectable by Western analysis.
HeLa cells containing a stably integrated tetracyclineinducible luciferase expression cassette were treated with Tet-Express according to the standard protocol and incubated overnight-in parallel with untreated cells from the same HeLa cell culture. The next day, lysates of treated and untreated cells were prepared and blotted with anti-luciferase and anti-actin antibodies. Basal expression is undetectable by Western analysis, but can be induced to very high levels in the presence of Tet-Express.
Figure 3. Vector formats for the Tet-Express Systems.
The Tet-Express Inducible Expression System (which contains the core pTRE3G vector) is also available in three other vector formats, for coexpressing two different genes of interest, or to monitor inducibility using induced coexpression of a red or green fluorescent protein.
1. Gossen, M. & Bujard, H. (1992) Proc. Natl. Acad. Sci. USA 89(12):5547-5551.
2. Gossen, M. et al. (1995) Science 268(5218):1766-1769.
3. Urlinger, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97(14):7963-7968.
4. Low, R., Heinz, N., Hampf, M., Bujard, H. & Gossen, M. (2010) BMC Biotechnology 10:81.
Each of the
Tet-Express Inducible Expression Systems
(Figure 3) is sold as a complete system that includes:
Tet-Express (25 rxns-includes Tet-Express Transactivator and Intensifier Reagent)
A tetracycline-inducible expression vector that expresses your gene of interest under the control of a P
promoter (the most advanced TRE promoter available)
A control vector (pTRE3G-Luc) that expresses luciferase under the control of a P
Both hygromycin and puromycin linear selection markers
Tet Approved FBS (50 ml)
Xfect transfection reagent (100 rxns)
Keyword : tet,dox,tetexpress,tetracycline,inducible
Tetracycline 유도 발현 시스템
양방향성 Tet 유도 발현 Vectors
Lenti-X Tet-On 3G Inducible Expression System
Linear 및 Plasmid 형 선택 Marker
Tet 시스템 발현 벡터
Tet System Approved FBS
Tet-Advanced Cell Lines
Tet-Inducible Dual Expression Systems
Tet-Off® & Tet-Off Advanced Cell Lines
Tet-On 3G Tatracycline 발현 유도 시스템
Tet-On Advanced & Tet-Off Advanced Tetracycline 유도발현 시스템
All-in-one vector 형태의 Tet-inducible system
Tetracycline Inducible Expression - Tet-One Systems