Difference between revisions of "Part:BBa J176017"
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Doxycycline-inducible promoter from Invitrogen vector pcDNA/FRT/TO, T-Rex system | Doxycycline-inducible promoter from Invitrogen vector pcDNA/FRT/TO, T-Rex system | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | * Chassis: mammalian cells | + | * Chassis: mammalian cells (Tet repressor +) |
* When used in cells without Tet, the promoter is constitutive (always on) | * When used in cells without Tet, the promoter is constitutive (always on) | ||
* Tetracycline is also an effective inducer. The levels of tetracycline in tet-treated cows is sufficient to cause a leaky off-state. Tet-free fetal bovine serum is highly recommended. | * Tetracycline is also an effective inducer. The levels of tetracycline in tet-treated cows is sufficient to cause a leaky off-state. Tet-free fetal bovine serum is highly recommended. | ||
<br> | <br> | ||
| − | '''CMVTetO2''' is a hybrid DNA element composed of the naturally occurring cytomegalovirus promoter (See CMV, <partinfo>BBa_J176027</partinfo>) and two tetracycline repressor binding sites/ operators (TetO x2). The TATA box motif stimulates formation of the transcription initiation complex at the promoter. The off-state of the promoter is maintained when the Tet repressor protein (expressed from a different gene) binds to the TetO x2 sites. When doxycycline or tetracycline is added to the cell culture medium, the chemical binds the Tet repressor | + | '''CMVTetO2''' is a hybrid DNA element composed of the naturally occurring cytomegalovirus promoter (See CMV, <partinfo>BBa_J176027</partinfo>) and two tetracycline repressor binding sites/ operators (TetO x2). The TATA box motif stimulates formation of the transcription initiation complex at the promoter. The off-state of the promoter is maintained when the Tet repressor protein (expressed from a different artificial gene) binds to the TetO x2 sites. When doxycycline or tetracycline is added to the cell culture medium, the chemical binds the Tet repressor and induces an allosteric change that decreases the repressor's affinity for the binding sites, and the promoter become active.<br> |
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| + | Transgenic mammalian cells that express the Tet repressor can be obtained from a commercial supplier (e.g., Invitrogen). | ||
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Latest revision as of 21:40, 18 February 2012
CMVTetO2
Doxycycline-inducible promoter from Invitrogen vector pcDNA/FRT/TO, T-Rex system
Usage and Biology
- Chassis: mammalian cells (Tet repressor +)
- When used in cells without Tet, the promoter is constitutive (always on)
- Tetracycline is also an effective inducer. The levels of tetracycline in tet-treated cows is sufficient to cause a leaky off-state. Tet-free fetal bovine serum is highly recommended.
CMVTetO2 is a hybrid DNA element composed of the naturally occurring cytomegalovirus promoter (See CMV, BBa_J176027) and two tetracycline repressor binding sites/ operators (TetO x2). The TATA box motif stimulates formation of the transcription initiation complex at the promoter. The off-state of the promoter is maintained when the Tet repressor protein (expressed from a different artificial gene) binds to the TetO x2 sites. When doxycycline or tetracycline is added to the cell culture medium, the chemical binds the Tet repressor and induces an allosteric change that decreases the repressor's affinity for the binding sites, and the promoter become active.
Transgenic mammalian cells that express the Tet repressor can be obtained from a commercial supplier (e.g., Invitrogen).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 605
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]