Difference between revisions of "Part:BBa K613022"
 
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===In vitro characterization===
 
  
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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 E37A P39K 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).
 
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WebLogo we obtained for the E37A P39K mutant:
 
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[[Image:EPFL2011_MITOMI_WebLogo_V36F.png|700px]]
 
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'''Reference:'''<p>
 
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Workman CT, Yin Y, Corcoran DL, Ideker T, Stormo GD, Benos PV.
 
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enoLOGOS: a versatile web tool for energy normalized sequence logos.
 
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Nucleic Acids Res. 2005 Jul 1;33:W389-92.</p>
 
  
 
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Latest revision as of 21:29, 21 September 2011

TetR E37A P39K mutant

This is a TetR mutant that carries the E37A P39K double mutation. Part of the EPFL2011 muTetR collection.

It has an altered tet operator sequence recognition compared to the wild-type tetR variant. And prefers the tetO-4C5G operator.



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]

Designed sequence (2 mutations in yellow):

MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVAQKTLYWHVKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCA

LLSHRDGAKVHLGTRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDSMPPLLRQAIELFD

HQGAEPAFLFGLELIICGLEKQLKCESGS*