Abstract
Annexin A3 (ANXA3) is a protein responsible for initiating angiogenesis and is associated with tumor growth and development. As a result, ANXA3 is used as a biomarker for several different kinds of cancer. When used in the context of profiling prostate cancer, drops in ANXA3 levels are associated with more severely disordered tissue biopsies. The purpose of this study was to express and isolate ANXA3 and purify the protein for future research. A strain of E.coli, C41(DE3), known for high protein yields when used with a pET system was transformed with the plasmid vector pET-28a(+) to produce ANXA3 when induced with varying levels of IPTG. The protein was purified under denaturing conditions and analyzed using a western blot. The results of this study show that ANXA3 yields the greatest protein concentration at three hours after induction; however, the SDS-page analysis indicated poor binding of the 6x-His tag to the nickel-based resin. The Western blot showed little to no antibody binding to the tagged target protein. Future experiments involving the moving of the 6x-His tag from the N to the C terminus and purifying under native conditions open up this experiment to potentially more efficient protein isolation.
Advisor
Morgan, William
Department
Biology
Recommended Citation
Gutierrez, Carson Jack, "Expression of ANXA3 in E. coli for the Development of Future Diagnostic Nanobodies" (2023). Senior Independent Study Theses. Paper 10484.
https://openworks.wooster.edu/independentstudy/10484
Disciplines
Biotechnology | Cancer Biology | Cell Biology
Keywords
Protein expression, ANXA3, prostate cancer, E. coli
Publication Date
2023
Degree Granted
Bachelor of Arts
Document Type
Senior Independent Study Thesis
© Copyright 2023 Carson Jack Gutierrez