Difference between revisions of "Part:BBa K176130"
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'''K176130'''
 
'''K176130'''
 
{| {{table}}
 
{| {{table}}
 +
{|class="sortable" style="border:3px solid black; width: 70%" align="center"
 
| align="center" style="background:#f0f0f0;"|'''T(hour)'''
 
| align="center" style="background:#f0f0f0;"|'''T(hour)'''
 
| align="center" style="background:#f0f0f0;"|'''OD600'''
 
| align="center" style="background:#f0f0f0;"|'''OD600'''
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| 3.5||0.558||267.6666667||479.6893668||1.00E-05||1917.453599
 
| 3.5||0.558||267.6666667||479.6893668||1.00E-05||1917.453599
 
|}
 
|}
−
  For the details of the definition of '''ST.FLU''' and the unit '''ustc_st1''', please click [[here]].  
+
  For the details of the definition of '''ST.FLU''' and the unit '''ustc_st1''', please click [[here]].
  
 
===plot===
 
===plot===

Revision as of 08:35, 20 October 2009

TetR repressible AHL->GFP Receiver (Low LuxR): pCon 0.04->luxR+pLux/Tet->GFP

AHL -> GFP Receiver

TetR repressible

Low LuxR level

K176129 with GFP.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 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 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

data

K176130

T(hour) OD600 FLU(arbitrary) FLU/OD(arbitrary) AHL ST.FLU/OD(ustc_st1)
0.5 0.140666667 0.832666667 5.91943128 0 23.66163521
0.5 0.146666667 0.747666667 5.097727273 1.00E-10 20.3770527
0.5 0.151 1.635 10.82781457 1.00E-09 43.28182664
0.5 0.144666667 5.035333333 34.80645161 1.00E-08 139.1311972
0.5 0.145 6.813666667 46.9908046 1.00E-07 187.8354903
0.5 0.147 7.474 50.84353741 1.00E-05 203.2359492
1.5 0.249666667 1.616333333 6.473965287 0 25.87826393
1.5 0.262 1.293666667 4.937659033 1.00E-10 19.73721483
1.5 0.266333333 1.912666667 7.181476846 1.00E-09 28.70638704
1.5 0.256 9.243 36.10546875 1.00E-08 144.3237349
1.5 0.248666667 26.90566667 108.1997319 1.00E-07 432.5048243
1.5 0.243 52.47433333 215.9437586 1.00E-05 863.1880664
2.5 0.407 3.353666667 8.23996724 0 32.93747148
2.5 0.414 2.235666667 5.400161031 1.00E-10 21.58596567
2.5 0.424 3.147333333 7.422955975 1.00E-09 29.67164718
2.5 0.41 15.49433333 37.79105691 1.00E-08 151.0615058
2.5 0.394333333 55.61766667 141.0422654 1.00E-07 563.7856874
2.5 0.39 139.1066667 356.6837607 1.00E-05 1425.765522
3.5 0.565666667 9.078 16.04832057 0 64.14966049
3.5 0.577333333 3.753 6.500577367 1.00E-10 25.98463991
3.5 0.607 5.145333333 8.476661175 1.00E-09 33.88360385
3.5 0.579666667 26.34833333 45.45428407 1.00E-08 181.6935846
3.5 0.557666667 98.57133333 176.7567244 1.00E-07 706.5464462
3.5 0.558 267.6666667 479.6893668 1.00E-05 1917.453599 
For the details of the definition of ST.FLU and the unit ustc_st1, please click here.

plot

K176130.jpg

protocol

AHLHybrid 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.