Part:BBa_K902090:Experience

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Applications of BBa_K902090

This part was tested electrochemically. CPRG was added to the solution, which is functional LacZ is produced, should be cleaved to CPR, which can be detected electrochemically using a potentiostat via an increase in current. CPR is able to oxidize at a specific voltage resulting in a change in current. The change in current can be correlated to the activity level of the promoter. Our results can be seen in figure 1 below.

Figure 1. Current production from the lacZ and uidA systems under the IPTG inducible promoter. Both samples were run with the same conditions and held at the oxidation potentials of their respective analytes.

In this figure we are measuring the change in current for this circuit, as well as for a circuit producing Beta-glucuronidase (UidA) under the same promoter. This enzyme cleaves a different substrate, PNPG, into PNP which also oxidizes, but at a different voltage to CPR. This allows for the two to be detected simultaneously. The differenece that can be seen in their induction levels is due to the fact that the analyte produced through the action of UidA produces a product that oxidizes to release two electrons while the LacZ product only releases one electron when oxidized. As the current we measure is this release of electrons, a similar amount of the two enzymes would results in the doubling of current for uidA that we saw. This graph thus indicates that our two circuits are working exactly as we expect them to!

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