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Team:Chiba/lux promoter
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==Abstract== | ==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 | + | 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> |
| - | The Plux inv was resistered in Biobrick number R0061.We've | + | |
==Experiments== | ==Experiments== | ||
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1,LBL | 1,LBL | ||
incubation for12 h at 37゜C. | incubation for12 h at 37゜C. | ||
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| - | + | ==Results== | |
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We confirmed GFP Fluorescence both AHL+ and -sample.<br> | We confirmed GFP Fluorescence both AHL+ and -sample.<br> | ||
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Latest revision as of 13:29, 24 January 2011
Contents |
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.
Experiments
1.Preparing Plux inv-GFP
Plasmid:R0061 and E0240
Strain:XL10-G
protocol
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.
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.
1,LBL
incubation for12 h at 37゜C.
Results
We confirmed GFP Fluorescence both AHL+ and -sample.
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)
