Difference between revisions of "Part:BBa K1895999:Experience"
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<img alt="Zinc Growth" src="https://static.igem.org/mediawiki/2016/f/f3/T--Newcastle--VR_Zinc_Growth.png"/>
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<img alt="Arabinose" src="https://static.igem.org/mediawiki/2016/0/01/T--Newcastle--VR_with_Arabinose.png"/>
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<p><figcaption>Figure 1. The minimum inhibitory concentration for <i>E. coli</i> growing in ZnSO<sub>4</sub>. The cells were left for 16 hours to grow in different zinc sulphate concentrations with LB broth and chlormaphenicol. OD600 was measured after this 16 hour growth period.</figcaption></p>
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<p><figcaption>From this experiment, we found that the minimum inhibitory concentration was 3mM of ZnSO<sub>4</sub> for <i>E. coli</i>. This concentration was then used in our second experiment to test whether our “Variable Resistor” (BBa_K1895999) construct was able to sequester zinc in the presence of arabinose.</figcaption></p>
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<p><figcaption>Figure 2. Growth curves of <i>E. coli</i>. Those growing with zinc were grown in 3mM of ZnSO4 with 0.5% arabinose and LB broth with chloramphenicol. Those growing without zinc were grown in 0.5% arabinose and LB broth with chloramphenicol. The OD600 was measured every five minutes, and the cultures were shaken in between the readings. The cells were grown for 16 hours. VR = Variable resistor BBa_K1895999, K1895100 = Our positive control BBa_K1895100, WT= Wild-type</figcaption> </p>
 
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Revision as of 21:47, 19 October 2016


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Applications of BBa_K1895999

User Reviews

UNIQd483df580f06accd-partinfo-00000000-QINU UNIQd483df580f06accd-partinfo-00000001-QINU

The overnight cultures of E. coli cells containing our BBa_K1895999 were diluted down to a starting optical density of around 0.05 at 600nm using LB broth with chloramphenicol. For characterisation of this device, we used a 96 well plate and used half of the plate to contain 3mM of ZnSO4 with 0.5% of arabinose and LB broth with chloramphenicol. The other half of the plate contained the 0.5% of arabinose with LB and chloramphenicol.The cells were then left to grow for 16 hours and the OD600 was measured every 5 minutes with orbital shaking ocuring inbetween.

Arabinose

Figure 2. Growth curves of E. coli. Those growing with zinc were grown in 3mM of ZnSO4 with 0.5% arabinose and LB broth with chloramphenicol. Those growing without zinc were grown in 0.5% arabinose and LB broth with chloramphenicol. The OD600 was measured every five minutes, and the cultures were shaken in between the readings. The cells were grown for 16 hours. VR = Variable resistor BBa_K1895999, K1895100 = Our positive control BBa_K1895100, WT= Wild-type

From our experiments, we can conclude that our ‘Variable Resistor’ (VR) construct does not grow in ZnSO4 when arabinose is present. The cells only grew when zinc was not present. Therefore, showing that our construct doesn’t sequester zinc when the SmtA protein is produced.