In 1992, Goseen et al. successfully constructed a tetracycline (Tet) eukaryotic gene regulatory expression system using prokaryotic gene regulatory elements. At present, this system has been widely used in the research of gene function and gene therapy.
The Tet-regulated expression system regulates the conformation of the regulatory protein by inducing a drug (such as Tet) to achieve the purpose of regulating the expression of the target protein.
The original Tet regulatory gene expression system was established based on the Tet resistance operon on the E. coli Tn10 transposon. The Tet repressor protein (TetR) and the Tet operator (TetO) are capable of specifically binding. When there is no Tet in the cell, Tet will bind to TetO, which blocks the expression of downstream resistance genes. When Tet is present, Tet changes the conformation of TetR, resulting in the separation of TetR from TetO and the expression of downstream resistance genes. Bacteria to obtain resistance
2, Tet-on control system
Using the characteristics of specific binding of TetR and TetO, various types of Tet regulatory systems have gradually developed. The most widely used is the Tet-on activated system.
The Tet-on system consists of a regulatory expression vector and a reaction expression vector.
The regulatory expression vector comprises a human cytomegalovirus early promoter (PhCMV) and a reverse tetracycline transcriptional activator (rtTA). Wherein rtTA is formed by fusion of a reverse transcriptional TetR (reverse TetR, rTetR) and a transcriptional activation region of the C-terminus of the herpes simplex virus (HSV) VP16 protein.
The reaction expression vector consists of a Tet response element (TRE), a minimal CMV promoter (Pmin CMV) and a gene of interest. Wherein TRE is 7 repeating TetO sequences.
Since PminCMV lacks an enhancer, the target gene is not expressed when rtTA is not bound to TRE; when rtTA binds to TRE, VP16 activates PminCMV to express the gene.
In the absence of Dox, rTetR cannot bind to TRE, resulting in inhibition of gene expression; and when Dox is present, rTetR can bind to TRE, thereby allowing expression of the gene of interest.
The Tet-on control system combines four characteristics of high efficiency, accuracy, safety and reversibility.
Virgin can provide you with adenovirus, lentivirus, adeno-associated virus and tetracycline-regulated expression vectors , as well as viral packaging services .
High Pressure Extension Tubings,high pressure PVC extension tube,Extension Tubings,extension tube medical
Anesthesia Medical Co., Ltd. , https://www.trustfulmedical.com