Team:Chiba/System 2/Result

From 2010.igem.org

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(1.Preparing Plux inv-GFP)
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===1.Preparing Plux inv-GFP===
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===1.Construction Plux inv-GFP===
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1.Construction of Plux inv-GFP
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Construction process is shown in '''Fig. 1'''.
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Construction process is shown in '''Fig. 1'''.Plux inv-GFP is constructed by conbining R0061 and E0240 and transformed strain of XL10-G.
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[[Image:Plux inv-GFP.png|none|frame|right|'''Fig. 1'''  Evaluation of LuxR Inverter]]
[[Image:Plux inv-GFP.png|none|frame|right|'''Fig. 1'''  Evaluation of LuxR Inverter]]

Revision as of 23:53, 27 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.Plux inv-GFP is constructed by conbining R0061 and E0240 and transformed strain of XL10-G.


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


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)