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From 2012.igem.org

Communication Bacteria<->Xenopus

Overview

Engineering the tadpole with AID system raised a question how will we deliver auxin to the embryo? One of the ideas was to use bacteria as a delivery machine in order to create a communication between two engineered organisms.

Steps

Our idea was to use previous biobricks from Imperial College 2011 BBa_K515100. Indeed, they managed to express in Escherichia coli the genes encoding the IAA-producing pathway from Pseudomonas savastanoi. Besides, we constructed a plasmid with a reporter (mRFP) as a control to monitor the auxin production.

Figure 1: Enginereed bacteria delivery to tadpole

Embryos were placed in medium containing MMR and DH5a bacteria with either BBa_K515100 or a reporter (mRFP), as shown on Figure 2.

Figure 2: Schematic representation of bacteria delivery to the embryos Figure 3: Fluorescence quantification The results show that, except for the control, all tadpoles are fluorescent. The Fig.2 shows also that the fluorescence occurs mainly in the intestinal tract. The death rate during for the experiments is close to 0%. We've performed the same using the Imperial College 2011 plasmid; the tadpoles didn't show any sign of auxin intolerance.