Difference between revisions of "Part:BBa K613017"
Line 4: Line 4:
 
This is a TetR mutant that carry the Y42F mutation.  
 
This is a TetR mutant that carry the Y42F mutation.  
 
Part of the <html> <a href="http://2011.igem.org/Team:EPF-Lausanne/Our_Project/TetR_mutants/muTetRs">EPFL2011 muTetR collection. </a> </html>
 
Part of the <html> <a href="http://2011.igem.org/Team:EPF-Lausanne/Our_Project/TetR_mutants/muTetRs">EPFL2011 muTetR collection. </a> </html>
 +
 +
===In vitro characterization===
 +
 +
Using the <html> <a href="http://2011.igem.org/Team:EPF-Lausanne/Our_Project/TetR_mutants/MITOMI_data">MITOMI</a></html> technique we determined the DNA binding landscape of the TetR Y42F mutant. To do so, first we designed and generated the library of double stranded DNA sequences that cover all possible single base substitution within the tetO operator sequence. Based on that library we measured the dissociation constants of the mutant to variable tetO-like sequences and determined the specificity of the mutant to the tet operator sequence (expressed as a PWM).
 +
 +
WebLogo we obtained for the Y42F mutant:
 +
 +
[[Image:EPFL_WebLogo_Y42F.png|700px]]
 +
 +
 +
'''Reference:'''<p>
 +
Workman CT, Yin Y, Corcoran DL, Ideker T, Stormo GD, Benos PV.
 +
enoLOGOS: a versatile web tool for energy normalized sequence logos.
 +
Nucleic Acids Res. 2005 Jul 1;33:W389-92.</p>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 21:32, 21 September 2011

TetR Y42F mutant

This is a TetR mutant that carry the Y42F mutation. Part of the EPFL2011 muTetR collection.

In vitro characterization

Using the MITOMI technique we determined the DNA binding landscape of the TetR Y42F mutant. To do so, first we designed and generated the library of double stranded DNA sequences that cover all possible single base substitution within the tetO operator sequence. Based on that library we measured the dissociation constants of the mutant to variable tetO-like sequences and determined the specificity of the mutant to the tet operator sequence (expressed as a PWM).

WebLogo we obtained for the Y42F mutant:

EPFL WebLogo Y42F.png


Reference:

Workman CT, Yin Y, Corcoran DL, Ideker T, Stormo GD, Benos PV. enoLOGOS: a versatile web tool for energy normalized sequence logos. Nucleic Acids Res. 2005 Jul 1;33:W389-92.

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
    COMPATIBLE WITH RFC[1000]