Part:BBa_K895007:Experience

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User Reviews

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••••• iGEM Dundee 2012

This part was seen work in practice. The device produces GFP in response to extracellular tetrathionate (Figure 1). The device is not activated by thiosulfate, which is the natural product of tetrathionate reduction (Figure 1).

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Results

E. coli MG1655 was transformed with BBa_K895007 grown aerobically overnight in LB medium. The culture was then used to inoculate 30 ml anaerobic cultures in LB only, LB + 0.4% (w/v) tetrathionate or LB + 0.4% (w/v) thiosulfate. Following 16 hours growth aliquotes were taken and analysed by SDS-PAGE and Western immunoblotting using an anti-GFP monoclonal.

This small scale aassay reveals that GFP was only expressed from the plasmid in the presence of tetrathionate. The device therefore works as a putative inflammation biosensor.

Figure 1: Production of GFP from BBa_K895007 is tetrathionate dependent. The RED ARROW points to the lane producing GFP.

References

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