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Team:Chiba/lux promoter
From 2010.igem.org
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===1.Preparing Plux inv-GFP=== | ===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. | 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. | ||
Revision as of 21:54, 27 October 2010
Contents |
Abstract
LuxR is an AHL-dependent activator. However to insert lux box between -35 and -10 and LuxR fuctions as LuxR-AHL inverter.
Someone resistered the Plux inverter in Biobrick.We've prepare Plux inv-GFP 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.
Fig. 1 Evaluation of LuxR Inverter
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.LB liquid incubate 37゜C,12 h
Fig. 2 work folw
Results
AHLのありおよびなしに関わらず蛍光が観察された。
We confirmed
Fig. 3
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)
