Part:BBa_K176083
cell density->PoPS: pCon 0.70->luxI-LVA+pCon 0.70->luxR+pLux/Tet->PoPS
Output PoPS = (Cell Density * pCon 0.70) + Input AHL
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 856
Illegal NheI site found at 879 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 698
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1834
MEASUREMENT
- K176026([[1]]) is a device which can response to different concentrations of AHL. Here, we use K176126 as a detector to examing the AHL concentrations produced by constitutive promoters + luxI.
constitutive promoters + luxI-LVA:
K176020( J23101->luxI-LVA);
K176021( J23107->luxI-LVA);
K176022( J23115->luxI-LVA);
K176023( J23109->luxI-LVA);
K176024( J23103->luxI-LVA);
measuring device:
K176217:( K176020 + K176026):( K176083 + GFP);
K176218:( K176021 + K176026):( K176084 + GFP);
K176219:( K176022 + K176026):( K176085 + GFP);
K176220:( K176023 + K176026):( K176086 + GFP);
K176221:( K176024 + K176026):( K176087 + GFP);
data
|
K176217 |
| T(min) | OD600 | FLU(arbitrary) | FLU/OD600(arbitrary) | ST.FLU/OD600(ustc_st1) | ST.FLU(ustc_st1) |
| 0 | 0.0875 | 100.2605 | 1147.557098 | 4587.109158 | 401.3720513 |
| 60 | 0.120666667 | 148.52 | 1229.380903 | 4914.181969 | 592.9779576 |
| 120 | 0.145 | 198.92 | 1370.07619 | 5476.580687 | 794.1041996 |
| 180 | 0.207666667 | 292.4516667 | 1404.89787 | 5615.772756 | 1166.208809 |
|
K176218 |
| T(min) | OD600 | FLU(arbitrary) | FLU/OD600(arbitrary) | ST.FLU/OD600(ustc_st1) | ST.FLU(ustc_st1) |
| 0 | 0.1035 | 75.11366667 | 721.4097924 | 2883.678268 | 298.4607008 |
| 60 | 0.150333333 | 123.0506667 | 808.6659075 | 3232.465553 | 485.9473215 |
| 120 | 0.1995 | 195.035 | 966.403576 | 3862.987472 | 770.6660008 |
| 180 | 0.308333333 | 351.1483333 | 1131.802978 | 4524.135501 | 1394.941779 |
|
K176219 |
| T(min) | OD600 | FLU(arbitrary) | FLU/OD600(arbitrary) | ST.FLU/OD600(ustc_st1) | ST.FLU(ustc_st1) |
| 0 | 0.2515 | 32.2705 | 125.5594805 | 501.8966322 | 126.227003 |
| 60 | 0.372833333 | 43.24566667 | 112.4353648 | 449.4358429 | 167.5646634 |
| 120 | 0.474833333 | 68.484 | 139.8281768 | 558.9326329 | 265.3998452 |
| 180 | 0.594833333 | 116.7865 | 192.791143 | 770.6405364 | 458.4026791 |
|
K176220 |
| T(min) | OD600 | FLU(arbitrary) | FLU/OD600(arbitrary) | ST.FLU/OD600(ustc_st1) | ST.FLU(ustc_st1) |
| 0 | 0.2485 | 7.723 | 32.1438897 | 128.4881869 | 31.92931443 |
| 60 | 0.4105 | 12.202 | 29.72814969 | 118.8317932 | 48.78045109 |
| 120 | 0.491666667 | 17.4355 | 36.20398381 | 144.7175273 | 71.15278427 |
| 180 | 0.611 | 31.26466667 | 51.84736322 | 207.2485239 | 126.6288481 |
|
K176221 |
| T(min) | OD600 | FLU(arbitrary) | FLU/OD600(arbitrary) | ST.FLU/OD600(ustc_st1) | ST.FLU(ustc_st1) |
| 0 | 0.219666667 | 11.49566667 | 52.33304675 | 209.1899378 | 45.95205634 |
| 60 | 0.3495 | 16.41366667 | 46.97848701 | 187.7862534 | 65.63129556 |
| 120 | 0.454833333 | 24.69433333 | 54.29412657 | 217.0289266 | 98.71199013 |
| 180 | 0.5765 | 43.4475 | 75.41615552 | 301.4596295 | 173.7914764 |
For the details of the definition of ST.FLU and the unit ustc_st1, please click here.
plot
protocol
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 1:100 into 3ml fresh medium and grown for 3hrs.
5. 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) every 30 minutes in the next 4hrs.
| None |