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Team:WPI-Worcester/Tour3
From 2014.igem.org
How do we plan to solve this problem?
Surface Expression of Proteins
We needed to express the antigen on the cell surface. We used the N-Terminus of BclA and the capsid protein of CAEV.
Ligate
Create the target sequence of BclA and CAEV p28. BclA is a membrane binding domain and CAEV-p28 is the capsid protein of CAEV that antibodies bind to.
Transform
Transform the target plasmid into e. coli which would then express CAEV-p28 on its surface.
Draw Blood
Take a sample of blood from the goat being diagnosed.
Add Bacteria
Mix the transformed e. coli with the sample of goat blood.
Agglutinate
If the disease is present in the goat being diagnosed, the antibodies for the disease in its blood will bind to the e. coli.
Diagnosis
The agglutinated diseased blood (+) will be visibly different from the non-diseased blood (-).
Agglutination as a Diagnostic
We based our agglutination assay off of the one from this paper.. Wells with dots are unagglutinated while wells with a thin film are agglutinated.
