Reporter

Part:BBa_K176112

Designed by: Danqian Liu, Hao Jiang   Group: iGEM09_USTC   (2009-09-07)
Revision as of 22:15, 20 October 2009 by Dqliu (Talk | contribs)

CI Inverter + GFP Reporter: VR9+pCon 0.15->cI-LVA+pCI->GFP

pCon 0.15 -> K176107

K176107:

High PoPS Input -> Low GFP Output

Low PoPS Input -> High GFP Output

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 35
    Illegal NheI site found at 58
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1728

MEASUREMENT

We measured the effect of inverter of Q04510 by test the response of these devices (these devices were transformed to  : 

K176112( J23115 + K176107);
K176113( J23109 + K176107);
K176114( J23103 + K176107);

OUTPUT: GFP

data

K1760112

COLONY T(h) OD600 FLU(arbitrary) ST.FLU(ustc_st1)
1 3 0.170333333 0.457333333 1.827871037
2 3 0.155333333 0.457666667 1.829203304
1 4 0.283 0.777 3.105515588
2 4 0.262666667 0.532 2.126298961
1 5 0.401333333 1.129333333 4.513722355
2 5 0.401333333 0.864333333 3.454569678
1 6 0.512 1.726 6.898481215
2 6 0.532333333 1.316666667 5.262456701

K1760113

COLONY T(h) OD600 FLU(arbitrary) ST.FLU(ustc_st1)
1 3 0.21 6.080333333 24.30189182
2 3 0.219 5.81 23.22142286
1 4 0.336666667 8.669666667 34.65094591
2 4 0.366 9.630333333 38.4905409
1 5 0.459 12.84733333 51.34825473
2 5 0.498 12.51266667 50.01065814
1 6 0.560666667 18.07833333 72.25552891
2 6 0.609 16.88866667 67.50066613

K1760114

COLONY T(h) OD600 FLU(arbitrary) ST.FLU(ustc_st1)
1 3 0.150333333 52.47366667 209.7268852
2 3 0.127 53.24233333 212.7990941
1 4 0.263 135.7333333 542.4993339
2 4 0.221 132.8766667 531.0818012
1 5 0.390333333 230.72 922.1422862
2 5 0.339333333 222.6233333 889.7815081
1 6 0.515 314.0566667 1255.222489
2 6 0.459666667 299.9233333 1198.734346 
For the details of the definition of ST.FLU and the unit ustc_st1, please click here.

plot

Figure 1-the relationship chart between ST.FLU and OD600. From the chart, we can see that the strength of the inverter is so strong that the expression of GFP is totally inhibited when the constitutive promoter is Pcon0.15

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.


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Parameters
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