Date of Award
Santa Clara University
Protein-protein interactions regulate key cellular functions and cell signaling pathways in the body; biological systems can be disrupted by protein interactions or lack of interactions that cause cell dysfunction and can lead to disease or illness. However, studying these interactions is complex and difficult to quantify. In this project, we examine the interaction of two putative cancer biomarkers, Bax and PDCDS, using the tetracycline repressor-based mammalian two-hybrid system (trM2H). This in vivo system engineers the cell as a biosensor using methods of DNA cloning and mammalian cell transfections that easily detects and quantities the interaction of two proteins by the production of GF P. The results of our experiments indicate that Bax and PDCDS do not directly interact but have a high affinity for forming individual homodimers. Although more experiments should be done to validate these claims, the genetic evidence we provide can dispute the proposed cell signaling pathway involving Bax and PDCDS with confidence. Success of our experiments also supports the trM2H system as a valid, versatile, and simple method of studying protein-protein interactions in vivo and with biological relevance.
Shimomura, Elyse and Perezalonso, Paulina, "The trM2H : detecting protein-protein interactions by engineering the cell as a biosensor" (2013). Bioengineering Senior Theses. 2.