Part:BBa_K1895999:Experience

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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.

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.

To see our results in more detail, check out the 2016 Newcastle University iGEM team's wiki