Part:BBa_K176128
TetR repressible AHL->GFP Receiver (Medium LuxR): pCon 0.15->luxR+pLux/Tet->GFP
AHL -> GFP Receiver
TetR repressible
Medium LuxR level
K176127 with GFP.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 985
Illegal BsaI.rc site found at 1729
MEASUREMENT
We mesured the hybrid promoter's response to AHL concentration through different devices. INPUT: AHL DEVICE: TetR repressible AHL->GFP Receiver: K176026 ( pCon 0.70->luxR+pLux/Tet->GFP ); K176126 ( pCon 0.36->luxR+pLux/Tet->GFP ); K176128 ( pCon 0.15->luxR+pLux/Tet->GFP ); K176130 (pCon 0.04->luxR+pLux/Tet->GFP ); OUTPUT: GFP
Model
To see the details of the model, click [http://2009.igem.org/Team:USTC/Modeling/Model-2 here] please.
data
K176128
T(hour) | OD600 | FLU(arbitrary) | FLU/OD(arbitrary) | AHL | ST.FLU/OD(ustc_st1) |
0.5 | 0.144 | 0.805 | 5.590277778 | 0 | 22.34591589 |
0.5 | 0.156 | 1.197333333 | 7.675213675 | 1.00E-10 | 30.67999231 |
0.5 | 0.160333333 | 4.548333333 | 28.36798337 | 1.00E-09 | 113.394825 |
0.5 | 0.16 | 7.007 | 43.79375 | 1.00E-08 | 175.0559619 |
0.5 | 0.161333333 | 8.152333333 | 50.53099174 | 1.00E-07 | 201.986616 |
0.5 | 0.157333333 | 8.976666667 | 57.05508475 | 1.00E-05 | 228.0652546 |
1.5 | 0.256666667 | 1.568 | 6.109090909 | 0 | 24.4197582 |
1.5 | 0.274 | 1.970333333 | 7.190997567 | 1.00E-10 | 28.74444405 |
1.5 | 0.282666667 | 9.054 | 32.03066038 | 1.00E-09 | 128.0355773 |
1.5 | 0.276333333 | 64.781 | 234.4306393 | 1.00E-08 | 937.0853393 |
1.5 | 0.274333333 | 89.16466667 | 325.0230863 | 1.00E-07 | 1299.208883 |
1.5 | 0.264333333 | 83.70166667 | 316.6519546 | 1.00E-05 | 1265.74711 |
2.5 | 0.356 | 2.742333333 | 7.703183521 | 0 | 30.79179566 |
2.5 | 0.391666667 | 3.256666667 | 8.314893617 | 1.00E-10 | 33.23697333 |
2.5 | 0.421333333 | 17.24066667 | 40.9193038 | 1.00E-09 | 163.5659903 |
2.5 | 0.396666667 | 169.93 | 428.394958 | 1.00E-08 | 1712.415389 |
2.5 | 0.391666667 | 271.61 | 693.4723404 | 1.00E-07 | 2772.004399 |
2.5 | 0.371 | 247.67 | 667.574124 | 1.00E-05 | 2668.481928 |
3.5 | 0.471666667 | 4.409 | 9.34770318 | 0 | 37.36540425 |
3.5 | 0.499 | 5.094 | 10.20841683 | 1.00E-10 | 40.80591931 |
3.5 | 0.557333333 | 28.99833333 | 52.03050239 | 1.00E-09 | 207.9805828 |
3.5 | 0.518 | 282.62 | 545.5984556 | 1.00E-08 | 2180.910803 |
3.5 | 0.514333333 | 463.9933333 | 902.1257291 | 1.00E-07 | 3606.050802 |
3.5 | 0.483333333 | 470.7433333 | 973.9517241 | 1.00E-05 | 3893.159548 |
For the details of the definition of ST.FLU and the unit ustc_st1, please click here.
plot
protocol
AHLHybrid promoter:BBa_K176026, BBa_K176126, BBa_K176128, BBa_K176130
1. Streak a plate of the strain which contain one of the parts listed in pSB1A3 .
2. Inoculate two 3ml cultures of supplemented M9 Medium and antibiotic(Ampicillin 0.1mg/ml) with single colony from the plate.
3. Cultures were grown in test tubes(BIO BASIC INC.12ml Polypropylene Round-bottom Culture Tubes With Graduations And Dual Cap Cat.No:TD444) for 16hrs at 37℃ with shaking at 200rpm.
4. Cultures were diluted 1:1000 to tubes of 3ml fresh medium and grown for 4.5hrs.
5. Stock concentration of the cognate AHL, 3-oxohexanoyl-homoserine is diluted and added to different tubes to yield different final concentrations (1E-5,1E-7,1E-8,1E-9,1E-10M).To ensure the same response time , the AHL should be added with a time interval of 2mins between tubes, so do the measurements procedure.
6. Measure the fluorescence(SHIMDZU SPECTROFLUOROPHOTOMETER RF-5301PC, 250ul quartz cell path length 10mm,501 nm excitation,514 nm emission,1.5nm slit width) and absorbance ((HITACHI UV-VIS spectrophotometer U-2810 ,200ul quartz cell path length 10mm,600nm,1.5 nm slit width) for the first time 30 minutes after adding AHL. Repeat measurement every 30 mins in the next 4hrs.
n/a | TetR repressible AHL->GFP Receiver (Medium LuxR): pCon 0.15->luxR+pLux/Tet->GFP |