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Team:Chiba/System 2/Result
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===<font size="5">Results</font>=== | ===<font size="5">Results</font>=== | ||
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| - | Pellet Photos is shown in'''Fig. 3''' | + | Pellet Photos is shown in '''Fig. 3'''.We confirmed GFP fluorescence both AHL+ and AHL-.So We're able to confirm LuxR repression. |
| - | [[Image:Plux inv results.png|none|right|frame|Fig. 3 | + | [[Image:Plux inv results.png|none|right|frame|Fig. 3 Pellet photos of the incubeted sample]] |
===Discussion=== | ===Discussion=== | ||
Revision as of 01:17, 28 October 2010
Version 2 :
Contents |
Abstract
LuxR is AHL-dependent activator.LuxR-AHL complex binds lux box, 20-bp sequence centered at position -42.5 from starting site and activates transcription.However lux box is inserted between -35 and -10,LuxR functions as AHL-inverter (Plux inv). Plux inv is resistered in Biobrick number R0061.We've prepared Plux inv-GFP and characterized about it.
Experiments
1.Construction Plux inv-GFP
Construction process is shown in Fig. 1.We constructed Plux inv-GFP conbining R0061 and E0240 and transformed by strain of XL10-G.
GFP fluorescence and sequence is confirmed.
2.Characterization Plux inv-GFP
strain:DH10B
sample
1,Plux inv-GFP and Plac-LuxR
2,Plux inv-GFP (Positive control)
3,Plux inv (Negative control)
add AHL1000 nM (AHL+)and
Each sample incubated for 12 h at 37゜C Spin-dawn and observed the pellet.
Results
Pellet Photos is shown in Fig. 3.We confirmed GFP fluorescence both AHL+ and AHL-.So We're able to confirm LuxR repression.
Discussion
We can't confirm LuxR repression.私たちは，LuxRの抑制を確認することができなかった。
原因として，私たちの実験レポータ遺伝子
Coxらの論文でもlux box を-35および-10の間に挿入しても，抑制効果がないことが示されている。
Reference
- Egland.K.A, and Greenberg.E.P, Conversion of the Vibrio Fischeri Transcriptional Activator,LuxR, to a Repressor, J. Bacteriol., 182, P.805-811 (2000)
- Cox.R.S.3rd, Surette.M.G, Elowitz.M.B, Programming gene expression with combinatorial promoters, Mol Syst Biol, 3 ,145 (2007)
