Difference between revisions of "Part:BBa K613013"
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− | ==In vitro characterization== | + | ===In vitro characterization=== |
[[Image:EPFL2011_MITOMI_WebLogo_V36F.png|700px]] | [[Image:EPFL2011_MITOMI_WebLogo_V36F.png|700px]] | ||
+ | |||
+ | {| {{table}} | ||
+ | | align="center" style="background:#f0f0f0;"|'''PO''' | ||
+ | | align="center" style="background:#f0f0f0;"|'''A''' | ||
+ | | align="center" style="background:#f0f0f0;"|'''T''' | ||
+ | | align="center" style="background:#f0f0f0;"|'''C''' | ||
+ | | align="center" style="background:#f0f0f0;"|'''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=== | ===In vivo characterization=== |
Revision as of 18:47, 21 September 2011
TetR V36F mutant
TetR with a point mutation V36F.
In vitro characterization
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.
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]