Difference between revisions of "Part:BBa K3861027"
Line 4: Line 4:
  
 
<html>
 
<html>
−
Composite part consisting of the Ptet-tetR tetracycline-inducible promoter <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K3861011">(Part:BBa_K3861011)</a> fused to GFP as reporter <a href="https://parts.igem.org/Part:BBa_E0840">(BBa_E0840)</a> derived from <i>E</i>. coli.
+
We have cloned our Salmonella codon harmonized GFP to a tetracycline-controlled transcriptional activation system (pTet-tetR) (BBa_K3861027{https://parts.igem.org/wiki/index.php?title=Part:BBa_K3861027}; [BBa_K3861011 + BBa_K3861019]). The tetR system is well characterized and may be the most used gene expression controlling promoter system to date.<i>1</i> There are two variants of this promoter-induction system, called Tet-On and Tet-Off.<I>2,3</i> 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.
 
</html>
 
</html>
  
Line 19: Line 19:
 
<partinfo>BBa_K3861027 parameters</partinfo>
 
<partinfo>BBa_K3861027 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
===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).

Revision as of 03:40, 22 October 2021


tetR-Ptet-GFP(STm codon harmonized)

We have cloned our Salmonella codon harmonized GFP to a tetracycline-controlled transcriptional activation system (pTet-tetR) (BBa_K3861027{https://parts.igem.org/wiki/index.php?title=Part:BBa_K3861027}; [BBa_K3861011 + BBa_K3861019]). 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).