Team:Cambridge/Bioluminescence/Vibrio Characterisation
From 2010.igem.org
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This page describes characterisation for part [http://partsregistry.org/Part:BBa_K325909 BBa K325909], the lux operon from Vibrio fischeri.
Description
![Cambridge-low.jpg](/wiki/images/4/40/Cambridge-low.jpg)
This page described the lux operon from Vibrio fischeri. To relieve LuxR control we placed Lux C, D, A, B, E under the pBad promoter.
Arabinose to light
This page describes the relationship between Arabinose concentration in the medium with light output. We used a [http://www.bmglabtech.com/products/microplate-reader/instruments.cfm?product_id=2 FLUOstar OPTIMA] microplate reader to quantify the light output. Protocol and plate reader settings used are given below.
Data
Data | Notes | Date Uploaded |
---|---|---|
Media:BBa_K325909ArabinosetoLight.xls | Raw data from experiment | 21/10/2010 |
H-NS mutants
It has been shown that the expression of the Vibrio fischeri lux operon when cloned into E. coli was repressed. This repression was linked to the nucleoid protein H-NS. To investigate this effect we cloned the operon into mutant E.coli cells in which the expression of the H-NS protein had been modified. We used a [http://www.bmglabtech.com/products/microplate-reader/instruments.cfm?product_id=2 FLUOstar OPTIMA] microplate reader to quantify the light output. Protocols and plate reader settings used are given below.
Data
![](/wiki/images/3/31/BBa_K325909timecourseK28.png)
![](/wiki/images/e/ed/BBa_K325909timecourseR28.png)
Data | Notes | Date Uploaded |
---|---|---|
Media:BBa_K325909Mutants.xls | Raw data from experiment | 21/10/2010 |
Compatibility
[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=cell Chassis:] Device has been shown to work in Top 10 (Invitrogen), [http://www.ecoliwiki.net/colipedia/index.php/BW25113 BW25113 DELTA H-NS::kan] and H-NS mutant JM 230 H-NS -205::tn10.
Plasmids: Device has been shown to work on <partinfo>pSB1C3</partinfo>
References
[http://www.jstor.org/stable/4449975 [1]:] J. Slock, (1995) Transformation Experiments Using Bioluminescence Genes of Vibrio fischeri,The American Biology Teacher, 57, 225-227.
[http://www.annualreviews.org/doi/pdf/10.1146/annurev.mi.42.100188.001055 [2]:] E.A. Meighen (1988) Enzymes and genes from the lux operons of bioluminescent bacteria, Annual Reviews in Microbiology 42, 151-176.
[http://www.annualreviews.org/doi/pdf/10.1146/annurev.ge.28.120194.001001 [3]:] E.A. Meighen, (1994) Genetics of bacterial bioluminescence, Annual Reviews of Genetics, 28, 117-139.
[http://onlinelibrary.wiley.com/doi/10.1002/%28SICI%291099-1271%28199807/08%2913:4%3C185::AID-BIO486%3E3.0.CO;2-U/abstract [4]:] S. Ulitzur, (1998) H-NS controls the transcription of three promoters of Vibrio fischeri lux cloned in Escherichia coli,Journal of Bioluminescence and Chemiluminescence, 13(4), 185-188.
[http://www.nature.com/nature/journal/v444/n7117/full/nature05283.html [5]:] R.T. Dame et al., (2006) Bacterial chromatin organization by H-NS protein unravelled using dual DNA manipulation,Nature, 444, 387-390.