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Xis-Mediated attL/R Inversion Assay

AttB/P flanking region inversion only counts for half of the reversible switch; Xis-mediated inversion is the other important part of the switch. Only when the two parts work, these bistable switch can be controlled to turn on and off downstream genes.

Fig. 1. RFP and qRT-PCR promoter analyses.

In case of the cytotoxicity of Xis and system chaos caused by constitutive expression of Xis, we put Xis CDS downstream of a L-arabinosed promoter, PBad. We constructed another plasmid called Reset and co-transform it with Int-6N to attest the inversion after Xis expresses.

Under the Arabinose concentration of 50mM, partial inversion from RFP to GFP was observed. Unfortunately, we don’t have enough time to tune the Xis expression relative to Int expression as what we do in the RBS modulation experiment.

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