Difference between revisions of "Team:Bielefeld-CeBiTec/Results/HeavyMetals"

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<!--Chrom bei niedrigeren Konzentrationen keinen signifikaten einfluss auf das System selbst, erst bei hoher Konzentration zeigt sich das CFPS nicht mehr in der lage ist ein Fluorescence signal zu erzeugen, zeigt, dass einen zu hohe konzentration das System zerstört. die Translations bzw Transskriptions ... sind nicht mehr in der Lage zu arbeiten, also keine
 
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<a href="https://static.igem.org/mediawiki/2015/9/99/Bielefeld-CeBiTec_Influence_of_chromium_on_the_cell_extract.jpeg" data-lightbox="heavymetals" data-title=" Influence of different chromium concentrations on our crude cell extract. Error bars represent the standard deviation of three biological replicates."><img src="https://static.igem.org/mediawiki/2015/9/99/Bielefeld-CeBiTec_Influence_of_chromium_on_the_cell_extract.jpeg" alt="Adjusting the detection limit"></a>
 
<a href="https://static.igem.org/mediawiki/2015/9/99/Bielefeld-CeBiTec_Influence_of_chromium_on_the_cell_extract.jpeg" data-lightbox="heavymetals" data-title=" Influence of different chromium concentrations on our crude cell extract. Error bars represent the standard deviation of three biological replicates."><img src="https://static.igem.org/mediawiki/2015/9/99/Bielefeld-CeBiTec_Influence_of_chromium_on_the_cell_extract.jpeg" alt="Adjusting the detection limit"></a>
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<a href="https://static.igem.org/mediawiki/2015/3/37/Bielefeld-CeBiTec_Influence_of_copper_on_the_cell_extract.jpeg" data-lightbox="heavymetals" data-title="Influence of different copper concentrations on our crude cell extract. Error bars represent the standard deviation of three biological replicates."><img src="https://static.igem.org/mediawiki/2015/3/37/Bielefeld-CeBiTec_Influence_of_copper_on_the_cell_extract.jpeg" alt="Adjusting the detection limit"></a>
 
<a href="https://static.igem.org/mediawiki/2015/3/37/Bielefeld-CeBiTec_Influence_of_copper_on_the_cell_extract.jpeg" data-lightbox="heavymetals" data-title="Influence of different copper concentrations on our crude cell extract. Error bars represent the standard deviation of three biological replicates."><img src="https://static.igem.org/mediawiki/2015/3/37/Bielefeld-CeBiTec_Influence_of_copper_on_the_cell_extract.jpeg" alt="Adjusting the detection limit"></a>

Revision as of 13:02, 14 September 2015

iGEM Bielefeld 2015


Heavy Metals

Zusammenfassung in ganz wenigen Worten.


The different sensors we worked with were characterized in vivo as well as in vitro.

We tested the influence of each heavy metal on our sensors in vivo Therefore we used heavy metal concentrations based on heavy metal occurrences measured all over the world.
Adjusting the detection limit
Influence of heavy metals on the growth of E.coli KRX shown is the standard deviation of three biological replicates. For induction concentrations of 20 µg/L lead, 60 µg/L mercury, 60 µg/L chromium, 80 µg/L nickel, 40 mg/L copper which represent ten times of the WHO guideline were used.


The tested heavy metal concentrations had no negative effect on E. colis growth. Moreover there is no significant difference between the curves with heavy metals and the controls. This first experiment showed us, in vivo characterization with these sensors under the tested heavy metal concentrations is possible. Most of our sensors were cultivated in the BioLector. Due to the accuracy of this device we could measure our sample in duplicates.

Arsenic

in vivo

Adjusting the detection limit
Time course of the induction of an arsenic biosensor with RFP for different arsenic concentrations in vivo. Error bars represent the standard deviation of three biological replicates.

in vitro

Adjusting the detection limit
Induction of arsenic sensor in vitro. For this experiment, a cell extract which already containes the arsenic repressor was used. Error bars represent the standard deviation of three biological replicates.

Chromium



in vivo



Our sensor for chromium detection consists of ChrB the repressor and the chromate specific promoter ChrP. The promoter is regulated by the ChrB, which binds Cr-ions. Behind the promoter is a sfGFP for detection of a fluorescence signal. In vivo we could show that the addition of different concentrations of chromium have different effects to transcription of sfGFP.

Adjusting the detection limit
Time course of the induction of a chromium biosensor with sfGFP for different chromium concentrations in vivo. The data are measured with BioLector and normalized on OD600. Error bars represent the standard deviation of two biological replicates.
Adjusting the detection limit
Fluorescence levels at three different stages of cultivation. Shown are levels after 60 minutes, 150 minutes and 650 minutes. Error bars represent the standard deviation of three biological replicates.

Our data lead to the conclusion that in a cell based system it is possible to detect chromium. In contrast to our expectations with higher chromium concentrations we got lower fluorescence levels. These observations needed further investigation.

in vitro

Adjusting the detection limit
Influence of different copper concentrations on our crude cell extract. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
Copper specific cell extract made from E. coli cells which have already expressed the activator before cell extract production. Induction of copper inducible promoter without T7 in front of the operator site with different copper concentrations. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
Copper specific cell extract made from E. coli cells which have already expressed the activator before cell extract production. Induction of copper inducible promoter without T7 in front of the operator site with different copper concentrations. Error bars represent the standard deviation of three biological replicates. Data are normalized on coppers influence to the cell extract.
Adjusting the detection limit
Copper specific cell extract made from E. coli cells which have already expressed the activator before cell extract production. Induction with different copper concentrations. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.

Lead

in vivo

Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.

Mercury

in vivo

Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.

in vitro

Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.

Nickel

in vivo

Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.
Adjusting the detection limit
TEXT. Error bars represent the standard deviation of three biological replicates.

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