Difference between revisions of "Part:BBa K1475005"
Line 20: Line 20:
 
<partinfo>BBa_K1475005 parameters</partinfo>
 
<partinfo>BBa_K1475005 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
===Characterization===
 +
We wanted to test if the Tet promoter could be fine-tuned, and what influence the LVA tag on TetR has on the expression.
 +
The ligation of the TetR(+LVA) construct with the pTet-GFP construct was cloned successfully and is found as [https://parts.igem.org/Part:BBa_K1475006 BBa_K1475006]
 +
 +
  
 
===References===
 
===References===

Revision as of 17:30, 17 October 2014

GFP controlled by TetR and pTet.

This composite part has GFP controlled by a TetR, p(TetR) expression system. The TetR repressor protein binds to the Tet promoter repressing expression. [1] The tet repressor protein is inhibited by [http://openwetware.org/wiki/ATc ATc (anhydrotetracycline)] or the analogue dox (doxycycline) hereby inducing the expression of GFP.[2]
Please note that this part contains the TetR repressor without the LVA rapid degradation tag, for a similar part containing TetR repressor with LVA tag please see Part:BBa_K1475006. The TetR+LVA bacis part can be found here: Part:BBa_C0040.


Usage and Biology

This part can be used to test the expression from the TetR p(TetR) system.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 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 1390


Characterization

We wanted to test if the Tet promoter could be fine-tuned, and what influence the LVA tag on TetR has on the expression. The ligation of the TetR(+LVA) construct with the pTet-GFP construct was cloned successfully and is found as BBa_K1475006


References

1. C. Krafft, et al.: Interaction of Tet Repressor with Operator DNA and with Tetracycline Studied by Infrared and Raman Spectroscopy. Biophysical Journal, Volume 74, Issue 1, January 1998, Pages 63–71. http://www.sciencedirect.com/science/article/pii/S0006349598777677
2. Tetsystems, 2008: Principles and Components Description. http://www.tetsystems.com/science-technology/principles-components
3. Cormack, B.P., Valdivia, R.H., and S. Falkow. FACS-optimized mutants of green fluorescent protein (GFP). Gene 173: 33-38 (1996). http://www.sciencedirect.com/science/article/pii/0378111995006850