Team:Queens Canada/Contribution
Contribution
Introduction
The Queens_Canada team hopes that our project can contribute to the success of future iGEM teams by making the use of the various aspect of synthetic biology easier to use and be more effective. In the past, iGEM teams have done many creative and helpful things to allow our team to have a better experience designing our project and our team hopes to do the same.
Our first contribution was to document the process of 3D printing our device. By analyzing the past iGEM teams and diagnostic teams like ourselves, we noticed that 3D printing can encompass a large portion of the project. Based on our experience, we noticed that designing parts and cases for 3D printing can take a lot of time, adjustments, and iterations before creating a good prototype. Our team experienced this firsthand. We printed many of the components that did not fit together which had to be remodeled, adjusted, or sanded. We hope that with our experience documented, future iGEM teams can make their own 3D printed components properly, quickly, and efficiently without having to redesign and reprint. Our 3D printing process can be found in the CAD Modeling Page.
Our next contribution, which was to aid in the detection of the spirochete, was unfortunately not completed due to time constraints. Initially, the team believed that using a chromogenic enzyme that can take a lot of substrates and produce a different coloured product would be successful. However, after some research, our team settled on alkaline phosphatase as it was highly compatible with the bioreceptor that we were attempting to express. Our team then looked at the parts currently available for alkaline phosphatase, and after analyzing recently published literature, we quickly realized that there was room for improvement. Our findings in the literature showed that a group had performed site-directed mutagenesis on alkaline phosphatase and had discovered a mutation in the active site which was able to increase the catalytic activity 40 times its basal rate in addition to discover various beneficial structural mutations. Our team adapted the catalytically optimized version of the alkaline phosphatase for use in the iGEM parts registry. Additionally, the researchers in the paper used E.Coli for protein expression and optimized the nucleotide sequence for E.Coli expression, making it optimal for use in the iGEM Parts Registry. Our team had planned to express and test this protein, our team did order the DNA sequence and clone it into an expression vector but ran out of time before we had a chance to express the protein. Although we didn’t get a chance to test the enzymatic activity firsthand, we wanted to share our findings with the iGEM community with the hopes that another team may be able to make use of it! The parts page can be found here:
http://parts.igem.org/Part:BBa_K3767001
