Team:Chiba/System 2/Result

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Revision as of 23:17, 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．Plux inv－GFPの作製クローニングの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.

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)

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 ===<font size="5">Abstract</font>===
 -----
-LuxR is AHL-dependent activator.BBa_R0061は-35および-10の間にlux box を含む配列である。継続的に下流遺伝子の転写を行う。しかしながら，LuxR-AHL複合体によって下流の遺伝子転写を抑制する。<br>
+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.<br><br><br>
-私たちは，Plux invの下流にRBSおよびGFP遺伝子を挿入したプラスミド (Plux inv-GFP) を作製し，抑制機能を確認した。<br>
-Biobrick number R0061 contains lux box between -35 and -10  and fuctions LuxR-AHL inverter.
-We've prepare Plux inv-GFP and characterized about it.
-<br><br><br>
 ===<font size="5">Experiments</font>===