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

From 2011.igem.org

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==E37A W43S T141A==
==E37A W43S T141A==
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<html> <a href=http://partsregistry.org/Part:BBa_K613015>BBa_K613015</a></html>
<html> <a href=http://partsregistry.org/Part:BBa_K613015>BBa_K613015</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 E37A  W43S T141A  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 W43S T141A mutant:
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[[Image:EPFL_WebLogo_EA37WS43TA41.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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{| {{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 ||0.298921 ||0.228558 ||0.136478 ||0.0565634
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|-
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|2 ||0.217725 ||0.298921 ||0.0365532 ||0.101569
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|-
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|3 ||0.486164 ||0.884721 ||0.298921 ||0.689301
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|-
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|4 ||0.381337 ||0.298921 ||1.2275 ||0.443129
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|-
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|5 ||0.298921 ||0.829256 ||1.48516 ||0.861923
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|-
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|6 ||1.52008 ||0.298921 ||1.20145 ||1.04388
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|-
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|7 ||1.08347 ||1.57658 ||0.298921 ||2.1751
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|-
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|8 ||0.298921 ||0.506289|| 0.723699 ||0.34977
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|-
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|9 ||0.0627581 ||0.298921|| 0 ||0.0173955
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|-
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|10 ||0.49991 ||0.298921 ||0.250601 ||0.869584
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|-
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|11 ||1.58199 ||1.5844 ||2.35356 ||0.298921
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|-
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|12 ||0.298921|| 1.88623|| 1.34104 ||1.46941
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|-
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|13 ||1.00088 ||0.298921 ||1.01931 ||1.57583
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|-
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|14 ||0.298921|| 0.56873 ||0.306484|| 1.91995
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|-
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|15 ||0.65822 ||1.06676 ||1.10609 ||0.298921
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|-
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|16 ||0.290169|| 0.067613 ||0.660542 ||0.298921
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|-
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|17 ||0.557743|| 0.160129 ||0.298921 ||0.134017
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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.
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===Position Weight Matrix===

Revision as of 01:55, 22 September 2011

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