Team:WPI-Worcester/Tour3
From 2014.igem.org
How do we plan to solve this problem?
![](https://static.igem.org/mediawiki/2014/c/c8/WPI_Team_Logo.png)
![](https://static.igem.org/mediawiki/2014/9/9a/WPI_Title.png)
Surface Expression of Proteins
![](https://static.igem.org/mediawiki/2014/a/a6/WPI_BCLA_NTD_Fig.jpg)
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.
![](https://static.igem.org/mediawiki/2014/2/2d/WPI_2014_Infographic1.png)
Transform
Transform the target plasmid into e. coli which would then express CAEV-p28 on its surface.
![](https://static.igem.org/mediawiki/2014/b/b2/WPI_2014_Infographic2.png)
Draw Blood
Take a sample of blood from the goat being diagnosed.
![](https://static.igem.org/mediawiki/2014/b/b1/WPI_2014_Infographic3.png)
Add Bacteria
Mix the transformed e. coli with the sample of goat blood.
![](https://static.igem.org/mediawiki/2014/f/fe/WPI_2014_Infographic4.png)
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.
![](https://static.igem.org/mediawiki/2014/b/b0/WPI_2014_Infographic5.png)
Diagnosis
The agglutinated diseased blood (+) will be visibly different from the non-diseased blood (-).
![](https://static.igem.org/mediawiki/2014/c/cd/WPI_2014_Infographic6.png)
Agglutination as a Diagnostic
![](https://static.igem.org/mediawiki/2014/3/36/WPI_How_to_Read_Agglutination.png)
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.