Team:Chiba/lux promoter

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Revision as of 21:44, 27 October 2010

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

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.LB liquid incubate 37゜C,12 h

Fig. 2 work folw

Results

AHLのありおよびなしに関わらず蛍光が観察された。
We confirmed

Fig. 3

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)

@@ Line 163: / Line 163: @@
 ===2.Characterization Plux inv-GFP===
-Plux inv-GFP およびLuxR generatorである Plac-LuxRをDH10B株に共形質転換した。コロニーをつついてLB液体培地(AHL0および1000 nM)で12 h培養し,
-スピンダウンしてペレットを観察した。
+Plasmid:Plux inv-GFP and Plac-LuxR (LuxR constitutive generator)<br>
-We cotransformded Plux inv-GFP and LuxR generator,Plac-LuxR in DH10B.
+strain:DH10B<br>
+protocol<br>
+We cotransformded Plux inv-GFP and LuxR Plac-LuxR in DH10B.LB liquid incubate 37゜C,12 h
 [[Image:lux inverter function.png|frame|center|Fig. 2 work folw]]