Summer Researcher with Dr. Dwight Higgin
University of Delaware
Zyairr’s Research Summary:
My summer research was at Willard Hall Education building at the University of Delaware, where I worked with Jazzlyn Jones on the cloning of the extracellular domain, one of the protein ABCA4.
ABCA4 is a part of the ATP binding cassette transporters
subfamily. It is found in the outer segment of the rod and cone photoreceptor cells. There are around 3000 which can lead to degenerative eye diseases such as Stargardt. This happens because ABCA4 is responsible for removing build up of toxic retinoid
compounds, and if there is a mutation the proteins will miss fold and now ABCA4 cannot correctly do its job, and those compounds will build up in the eye.
The research required the protein to be cloned, this was done using a polymerase chain reaction. This targets the desired sequence and amplifies it. The PCR amplified ECD1 undergoes a clean up then restriction digest and a gel purification. The ECD1 sequence is placed within the pET30 vector by restriction digest and ligation. During transformation the DNA was introduced to DH5 alpha E.coli cells for propagation. DNA from DH5 alpha was purified. A restriction digest was performed on the DNA to determine if it was recombinant. Five
samples were run on 0.8% agarose gel, where all resulted in positive. The next step would have been to transform the samples into BL12 for protein production.
This research gave me lab experience allowing me to learn how to use various lab equipment related to my intended major and how to study material such as DNA and what the results mean. I learned how to write and perform proper scientific procedures. I also learned how to properly create and present presentations and posters, along with how to categorize and organize the research done.