Team:Chiba/lux promoter

From 2010.igem.org

(Difference between revisions)
(Results)
(2.Characterization Plux inv-GFP)
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Plasmid:Plux inv-GFP and Plac-LuxR (LuxR constitutive generator)<br>
Plasmid:Plux inv-GFP and Plac-LuxR (LuxR constitutive generator)<br>
strain:DH10B<br>
strain:DH10B<br>
 +
protocol<br>  
protocol<br>  
We cotransformded Plux inv-GFP and LuxR Plac-LuxR in DH10B.
We cotransformded Plux inv-GFP and LuxR Plac-LuxR in DH10B.
-
LB liquid incubate 37゜C,12 h
+
1,LBL
 +
incubation for12 h at 37゜C
 +
Plux inv-GFPおよびLuxR generatorであるPlac-LuxR (PSB1A3)を共形質転換した。
 +
 
 +
得られたコロニーをpre cultureしたあと,AHL100 nM(AHL+)
==Results==
==Results==

Revision as of 22:07, 27 October 2010




Contents

Abstract

LuxR is an AHL-dependent activator. However to insert lux box between -35 and -10 and LuxR fuctions as LuxR-AHL inverter. Someone resistered the Plux inverter in Biobrick.We've prepare Plux inv-GFP and characterized about it.

Experiments

1.Preparing Plux inv-GFP

Plasmid:R0061 and E0240
Strain:XL10-G
protocol
Preparing process of Plux inv-GFP is shown in Fig. 1.We transformed preparing Plux inv-GFP in XL10-G and confirmed the expression GFP Fluorescence.The sequence is also conformed.

Fig. 1  Evaluation of LuxR Inverter

2.Characterization Plux inv-GFP

Plasmid:Plux inv-GFP and Plac-LuxR (LuxR constitutive generator)
strain:DH10B

protocol
We cotransformded Plux inv-GFP and LuxR Plac-LuxR in DH10B. 1,LBL incubation for12 h at 37゜C Plux inv-GFPおよびLuxR generatorであるPlac-LuxR (PSB1A3)を共形質転換した。

得られたコロニーをpre cultureしたあと,AHL100 nM(AHL+)

Results

AHLの有無に関わらず蛍光が観察された。
We confirmed GFP Fluorescence both AHL+ and -sample.

Fig. 3

Reference

  1. 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)
  2. Cox.R.S.3rd, Surette.M.G, Elowitz.M.B, Programming gene expression with combinatorial promoters, Mol Syst Biol, 3 ,145 (2007)