Team:Chiba/System 2/Result

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Revision as of 23:21, 27 October 2010

System 2 :

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.Preparing Plux inv-GFP

1．Construction of Plux inv-GFP

Construction process is shown in Fig. 1.

Fig. 1 　Evaluation of LuxR Inverter

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。

Fig. 2 assay protocol

Results

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 355: / Line 355: @@
 ===1.Preparing Plux inv-GFP===
-．Plux inv－GFPの作製
+．Construction of Plux inv-GFP
-クローニングのprocessをFig. 1に示す。作製したPlux inv-GFPを形質転換した株は緑色蛍光を示した。また，sequenceも正しいことを確認した。
-Preparing Plux inv-GFP process is shown in '''Fig. 1'''.
+Construction process is shown in '''Fig. 1'''.
 [[Image:Plux inv-GFP.png|none|frame|right|'''Fig. 1''' 　Evaluation of LuxR Inverter]]