Difference between revisions of "Part:BBa K613013"

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==In vitro characterization==
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===In vitro characterization===
  
 
[[Image:EPFL2011_MITOMI_WebLogo_V36F.png|700px]]
 
[[Image:EPFL2011_MITOMI_WebLogo_V36F.png|700px]]
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{| {{table}}
 +
| 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'''
 +
| 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||0.164072||0.519334||-0.0468541||0.22966
 +
|-
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| 2||0.642569||0.164072||0.188965||0.943314
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|-
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| 3||1.04481||0.558693||0.164072||0.848264
 +
|-
 +
| 4||0.959646||0.164072||2.17536||0.718959
 +
|-
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| 5||0.164072||1.19894||1.79469||1.56685
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|-
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| 6||1.46463||0.164072||1.74384||1.54845
 +
|-
 +
| 7||0.966397||1.07557||0.164072||1.72883
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|-
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| 8||0.164072||0.504312||0.755195||0.148902
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|-
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| 9||0.0876645||0.164072||0||0.0156484
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|-
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| 10||0.289722||0.164072||0.00707563||0.843474
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|-
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| 11||1.77568||1.3447||2.34804||0.164072
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|-
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| 12||0.164072||1.72354||1.62782||1.49176
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|-
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| 13||0.877518||0.164072||1.30879||1.9052
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|-
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| 14||0.164072||0.544642||0.3387||1.7537
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|-
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| 15||0.540091||0.8821||1.0861||0.164072
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|-
 +
| 16||-0.224147||0.280358||0.442769||0.164072
 +
|-
 +
| 17||0.31238||-0.0651059||0.164072||-0.303844
 +
|}
  
 
===In vivo characterization===
 
===In vivo characterization===

Revision as of 18:47, 21 September 2011

TetR V36F mutant

TetR with a point mutation V36F.


In vitro characterization

EPFL2011 MITOMI WebLogo V36F.png

PO A T C G
1 0.164072 0.519334 -0.0468541 0.22966
2 0.642569 0.164072 0.188965 0.943314
3 1.04481 0.558693 0.164072 0.848264
4 0.959646 0.164072 2.17536 0.718959
5 0.164072 1.19894 1.79469 1.56685
6 1.46463 0.164072 1.74384 1.54845
7 0.966397 1.07557 0.164072 1.72883
8 0.164072 0.504312 0.755195 0.148902
9 0.0876645 0.164072 0 0.0156484
10 0.289722 0.164072 0.00707563 0.843474
11 1.77568 1.3447 2.34804 0.164072
12 0.164072 1.72354 1.62782 1.49176
13 0.877518 0.164072 1.30879 1.9052
14 0.164072 0.544642 0.3387 1.7537
15 0.540091 0.8821 1.0861 0.164072
16 -0.224147 0.280358 0.442769 0.164072
17 0.31238 -0.0651059 0.164072 -0.303844

In vivo characterization

This TetR mutant was characterized in vivo by putting it into pSB3K1 under a constitutive promoter (J23116). This plasmid was cotransformed with J61002 harbouring RFP under pTet promoter (B0040) in DH5alpha cells. Cells were grown in a medium containing Kanamycin & Amplicillin plus different concentrations of ATC, ranging from 0 to 2000 ng/mL. OD600 absorbence and RFP fluorescence were measured every 10 minutes during 12 hours on a platereader machine.


Induction curves


Fluorescence measurements (RFUs) were normalized by OD600 values.

EPFL TetR-V36F-doseresponse.png


Dose-response curve


Fluorescence measurements (RFUs) were normalized by OD600 values. For each ATC concentration, we estimated the steady-state fluorescence expression by averaging the measurements over the last hour.

EPFL TetR-V36F-induction.png

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]