Difference between revisions of "Part:BBa K3861027"

 
 
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We have cloned our Salmonella codon harmonized GFP (BBa_K3861019) to a tetracycline-controlled transcriptional activation system (pTet-tetR, BBa_K3861011 ). The tetR system is well characterized and may be the most used gene expression controlling promoter system to date.<sup>1</sup> There are two variants of this promoter-induction system, called Tet-On and Tet-Off.<sup>2,3</sup> We use the Tet-On variant that selectively activates expression in the presence of tetracyclines. Important for our needs is the fact that tetracycline (or Anhydro-tetracycline) can passively penetrate bacterial membranes and no active transportes systems is crucial for this inducible expression system.
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===Reference===
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1. Gossen, M. & Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. U. S. A. 89, 5547–5551 (1992).
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2. Baron, U. & Bujard, H. Tet repressor-based system for regulated gene expression in eukaryotic cells: principles and advances. Methods Enzymol. 327, 401–421 (2000).
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3. Berens, C. & Hillen, W. Gene regulation by tetracyclines. Constraints of resistance regulation in bacteria shape TetR for application in eukaryotes. Eur. J. Biochem. 270, 3109–3121 (2003).

Latest revision as of 03:49, 22 October 2021


tetR-Ptet-GFP(STm codon harmonized)

We have cloned our Salmonella codon harmonized GFP (BBa_K3861019) to a tetracycline-controlled transcriptional activation system (pTet-tetR, BBa_K3861011 ). The tetR system is well characterized and may be the most used gene expression controlling promoter system to date.1 There are two variants of this promoter-induction system, called Tet-On and Tet-Off.2,3 We use the Tet-On variant that selectively activates expression in the presence of tetracyclines. Important for our needs is the fact that tetracycline (or Anhydro-tetracycline) can passively penetrate bacterial membranes and no active transportes systems is crucial for this inducible expression system.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1382


Reference

1. Gossen, M. & Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. U. S. A. 89, 5547–5551 (1992). 2. Baron, U. & Bujard, H. Tet repressor-based system for regulated gene expression in eukaryotic cells: principles and advances. Methods Enzymol. 327, 401–421 (2000). 3. Berens, C. & Hillen, W. Gene regulation by tetracyclines. Constraints of resistance regulation in bacteria shape TetR for application in eukaryotes. Eur. J. Biochem. 270, 3109–3121 (2003).