View source for Part:BBa K176126

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<partinfo>BBa_K176126 short</partinfo>

AHL -> GFP Receiver

TetR repressible

High LuxR level

[[Part:BBa_K176125|K176125]] with GFP.

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===Usage and Biology===

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<span class='h3bb'>Sequence and Features</span>
<partinfo>BBa_K176126 SequenceAndFeatures</partinfo>

=='''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===
'''K176126'''
{| {{table}}
{|class="sortable" style="border:3px solid black; width: 70%" align="center" 
| align="center" style="background:#f0f0f0;"|'''T(hour)'''
| align="center" style="background:#f0f0f0;"|'''OD600'''
| align="center" style="background:#f0f0f0;"|'''FLU(arbitrary)'''
| align="center" style="background:#f0f0f0;"|'''FLU/OD(arbitrary)'''
| align="center" style="background:#f0f0f0;"|'''AHL'''
| align="center" style="background:#f0f0f0;"|'''ST.FLU/OD(ustc_st1)'''
|-
| 0.5||0.147||0.724||4.925170068||0||19.68729291
|-
| 0.5||0.149666667||2.844||19.00222717||1.00E-10||75.95725775
|-
| 0.5||0.157||9.368666667||59.67303609||1.00E-09||238.529944
|-
| 0.5||0.152||11.27633333||74.18640351||1.00E-08||296.5439641
|-
| 0.5||0.149333333||11.57933333||77.54017857||1.00E-07||309.9499483
|-
| 0.5||0.147||12.41233333||84.43764172||1.00E-05||337.5210526
|-
| 1.5||0.268||1.283666667||4.789800995||0||19.14618457
|-
| 1.5||0.276666667||3.247666667||11.73855422||1.00E-10||46.9223097
|-
| 1.5||0.282666667||38.829||137.3667453||1.00E-09||549.0935975
|-
| 1.5||0.270333333||94.57733333||349.8545006||1.00E-08||1398.467045
|-
| 1.5||0.255666667||92.93366667||363.4954368||1.00E-07||1452.993711
|-
| 1.5||0.232||75.93966667||327.3261494||1.00E-05||1308.414876
|-
| 2.5||0.381666667||2.555||6.694323144||0||26.75909639
|-
| 2.5||0.439||5.467||12.45330296||1.00E-10||49.77936188
|-
| 2.5||0.437333333||92.855||212.3208841||1.00E-09||848.7064162
|-
| 2.5||0.409333333||227.38||555.4885993||1.00E-08||2220.444495
|-
| 2.5||0.382||222.79||583.2198953||1.00E-07||2331.294301
|-
| 2.5||0.299333333||163.65||546.714922||1.00E-05||2185.373634
|-
| 3.5||0.584||4.336||7.424657534||0||29.67844879
|-
| 3.5||0.611||7.840333333||12.83196945||1.00E-10||51.29299856
|-
| 3.5||0.599333333||154.5333333||257.8420467||1.00E-09||1030.667333
|-
| 3.5||0.574333333||412.9266667||718.9669182||1.00E-08||2873.913412
|-
| 3.5||0.518333333||401.8466667||775.266881||1.00E-07||3098.960231
|-
| 3.5||0.381666667||272.7966667||714.7510917||1.00E-05||2857.061565
|}

 For the details of the definition of '''ST.FLU''' and the unit '''ustc_st1''', please click [[here]]. 

===plot===

[[Image:126.jpg|center]]

===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.



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===Functional Parameters===
<partinfo>BBa_K176126 parameters</partinfo>
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