Difference between revisions of "Part:BBa K895007:Experience"

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===User Reviews===
 
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<I>Username</I>
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<I>iGEM Dundee 2012</I>
 
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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===
 
===Results===
 
<i>E. coli</i> MG1655 was transformed with <partinfo>BBa_K895007</partinfo> 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.
 
<i>E. coli</i> MG1655 was transformed with <partinfo>BBa_K895007</partinfo> 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.

Revision as of 08:55, 29 September 2012

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K895007

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).

UNIQff452536871c17fb-partinfo-00000002-QINU

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.

Dundee Biosensor Figure5.jpg

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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