Team:EPF-Lausanne/Our Project/TetR mutants/MITOMI data

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<html> <a href=http://partsregistry.org/Part:BBa_K613019>BBa_K613019</a></html>
<html> <a href=http://partsregistry.org/Part:BBa_K613019>BBa_K613019</a></html>
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===In vitro characterization===
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Using the <html> <a href="https://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). For the Y42F mutant we observed the decrease of the specificity compared to the wild-type tetR sequence.
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WebLogo we obtained for the Y42F mutant:
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[[File:EPFL2011_P39QY42M_WebLogo.png|700px]]
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====Position Weight Matrix====
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''PO'''
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| align="center" style="background:#f0f0f0;"|'''A'''
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| align="center" style="background:#f0f0f0;"|'''T'''
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| align="center" style="background:#f0f0f0;"|'''C'''
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| align="center" style="background:#f0f0f0;"|'''G'''
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|-
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| 1||0.455977||0.0513378||0.0469535||0.0942038
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|-
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| 2||0||0.455977||0.0808971||0.0550434
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|-
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| 3||0.024486||0||0.455977||0
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|-
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| 4||0||0.455977||0||0.175581
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|-
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| 5||0.455977||0||0.0549319||0
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|-
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| 6||0.162363||0.455977||0.0897636||0.0689181
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|-
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| 7||0||0.02975||0.455977||0.0170034
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|-
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| 8||0.455977||0||0||0.0975597
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|-
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| 9||0.134079||0.455977||0||0
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|-
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| 10||0.486152||0.455977||0.0220095||0
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|-
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| 11||0||0.0246322||0.0847586||0.455977
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|-
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| 12||0.455977||0||0||0.0321177
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|-
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| 13||0.20916||0.455977||0||0.0482279
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|-
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| 14||0.455977||0||0.0837475||0.141681
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|-
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| 15||0||0.107215||0||0.455977
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|-
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| 16||0||0||0.269667||0.455977
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|-
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| 17||0.0634094||0||0.455977||0.0582705
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|}
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Each row represents the changes in binding energy,  ΔΔG, compared to the reference sequence upon the substitution to the indicated nucleotide at certain position within the target DNA element. Values are indicated in kcal/mol.
==P39Q Y42M L197S==
==P39Q Y42M L197S==

Revision as of 02:13, 22 September 2011

Retrieved from "http://2011.igem.org/Team:EPF-Lausanne/Our_Project/TetR_mutants/MITOMI_data"