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Example of a T-REx system controlling the expression of shRNA Tetracycline-controlled transcriptional activation is a method of inducible gene expression where transcription is reversibly turned on or off in the presence of the antibiotic tetracycline or one of its derivatives (e.g. doxycycline ).
The overall structure of TetR can be broken down into two DNA-binding domains (one per monomer) and a regulatory core, which is responsible for tetracycline recognition and dimerization. TetR dimerizes by making hydrophobic contacts within the regulatory core. There is a binding cavity for tetracycline in the outer helices of the regulatory domain.
The use of bacteria-derived tetracycline-inducible system permitting the switching on or off (Tet-On/Tet-Off system) [36] Targeted mutations by knock in gene and knock out sequence by using Cre-Lox recombination system [37] Introduction of retro viral mutations [38] Introduction of chemically induced mutations
Members of the tetracycline class of antibiotics are often used as research reagents in in vitro and in vivo biomedical research experiments involving bacteria as well in experiments in eukaryotic cells and organisms with inducible protein expression systems using tetracycline-controlled transcriptional activation. [61]
The GAL4/UAS system is an example of both an inducible and repressible system. Gal4 binds an upstream activation sequence (UAS) to activate the transcription of the GAL1/GAL7/GAL10 cassette. On the other hand, a MIG1 response to the presence of glucose can inhibit GAL4 and therefore stop the expression of the GAL1/GAL7/GAL10 cassette. [42]
Tetracycline hydrochloride is available as yellow crystalline powder. Since tetracycline is absorbed into bone, it is used as a marker of bone growth for biopsies in humans. Tetracycline labeling is used to determine the amount of bone growth within a certain period of time, usually a period around 21 days.
Doxycycline is more stable than tetracycline for this purpose. [152] At subantimicrobial doses, doxycycline is an inhibitor of matrix metalloproteases, and has been used in various experimental systems for this purpose, such as for recalcitrant recurrent corneal erosions. [153]
In some systems, however, the protein may be expressed constitutively. Escherichia coli is commonly used as the host for protein production, but other cell types may also be used. An example of the use of expression vector is the production of insulin, which is used for medical treatments of diabetes.