Team:Chiba/lux promoter

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==Abstract==

LuxR is an AHL-dependent activator.LuxR-AHL complex bind lux box,a 20-bp sequence and activate transcription.  However the lux box is inserted between -35 and -10 and LuxR functions as LuxR-AHL inverter (Plux inv) .The Plux inv was resistered in Biobrick number R0061.We've constructed Plux inv-GFP combinig R0061and E0240 and characterized about it.<br>

==Experiments==
===1.Preparing Plux inv-GFP===

Plasmid:R0061 and E0240<br>
Strain:XL10-G<br>
protocol<br>
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.

[[Image:Plux inv-GFP.png|none|frame|right|'''Fig. 1''' 　Evaluation of LuxR Inverter]]

===2.Characterization Plux inv-GFP===

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

protocol<br> 
We cotransformded Plux inv-GFP and LuxR Plac-LuxR in DH10B.
1,LBL
incubation for12 h at 37゜C.

==Results==

We confirmed GFP Fluorescence both AHL+ and -sample.<br>
<br>
[[Image:Plux inv results.png|frame|none|Fig. 3 ]]

==discussion==
私たちの系では，LuxRの抑制を確認することができなかった。

==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)