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Selecting Circuit

We have explored the potential to use a series of antibiotic resistance genes as quantitative reporter, which their expression levels are converted into different growth rates of chassis organisms. In order to make the characterization data reusable, we have substituted pPsi promoter to pLac. pLac is a more characterized promoter than D-psicose induced one.

We designed a series of experiments to characterize the function of selecting circuit and to decide which antibiotic is suitable for our system. Take selecting circuit-KanR for example, we put the same quantity of bacteria with selecting circuit into a 96-well plate, added different concentration of Kan and IPTG into the medium and measured the OD600 during 5h. The results were shown as below.

             
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Figure 3-7 Interaction of AB and AB resistance proteins

As for selecting circuit-AmpR, only a little expression of AmpR can give bacteria extraordinary resistance. Although we didn’t add any inducer to the medium, the bacteria grown merrily in the ocean of Amp. Selecting circuit-SmR is the same. There is no doubt that Amp and Smr are not competent to screening. We chose KanR to construct our device D.

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