Crosstalk Confirmation Assay

Introduction

Our goal in this project is to construct a system to suppress crosstalk by 3OC12HSL-LasR complex on lux promoter. We thought that we could prove that our system precisely works only after we obtain data of crosstalk happening by ourselves. Therefore, we confirmed that crosstalk is really happening by the following assay.

Summary of the experiment

Our purpose is to confirm 3OC12HSL-LasR complex really activates lux promoter. We prepared four plasmids shown in below (Fig.1). We checked what would happen when we add intercellular molecules 3OC6HSL and 3OC12HSL.

We prepared twelve conditions as follow.

• A-1) Culture containing Ptrc-lasR and Plas-GFP cell with 3OC6HSL induction
• A-2) Culture containing Ptrc-lasR and Plas-GFP cell with 3OC12HSL induction
• A-3) Culture containing PtrclasR and Plas-GFP cell with DMSO ( no induction)

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• B-1) Culture containing Ptrc-lasR and Plux-GFP cell with 3OC6HSL induction