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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.

Figure 3-7 Interaction of AB and AB resistance proteins

When the concentration of IPTG locates in a certain range (below 1.5mM), the final OD600 of bacteria will decrease as the concentration of Kan increase. It implies that the bacteria have a high possibility of survival if the expression level of KanR is high, because a higher IPTG concentration can induce more expression of KanR. Moreover, the bacteria which expressed more KanR survived more.

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