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Thesis
Methodologies for protein expression and purification of VanU and AsnB proteins
Master of Science (M.S.), Drexel University
Aug 2023
DOI:
https://doi.org/10.17918/00001824
Abstract
Protein expression and purification are critical steps for studying the structure, function, and mechanisms of proteins. This thesis focuses on developing methodologies to efficiently express and purify two key proteins, VanU and AsnB, to enhance our understanding of their mechanisms and potential applications in biochemistry. The VanU protein, which is part of the vancomycin resistance cluster, regulates the expression of vancomycin resistance genes. Understanding the expression and purification of VanU is vital for uncovering its molecular activity and investigating its potential as a target for new antibiotics. This study explores different expression systems, vectors, primers, culture conditions, and purification strategies to obtain highly pure and active samples of the VanU protein. The AsnB protein plays a critical role in peptidoglycan precursor amidation in Clostridioides difficile (C. diff.) when vancomycin is present. Elucidating the expression and purification of AsnB is essential for studying its molecular mechanism and exploring its potential in treating C. diff. colitis. This research evaluates different expression vectors, host strains, and growth conditions to improve the yield and purity of AsnB protein preparations. Overall, this thesis outlines methodologies that enable the efficient expression and purification of VanU and AsnB proteins. The optimization of expression systems, culture conditions, and purification techniques allows researchers to obtain enough pure proteins for detailed biochemical and biophysical analyses. Furthermore, these methodologies have broader implications in the field of protein expression and purification, laying the groundwork for future investigations into other proteins of interest and facilitating advancements in scientific and industrial applications.
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Details
- Title
- Methodologies for protein expression and purification of VanU and AsnB proteins
- Creators
- Yagni Keyur Shah
- Contributors
- Paul McGonigle (Advisor)Patrick J. Loll (Advisor)
- Awarding Institution
- Drexel University
- Degree Awarded
- Master of Science (M.S.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Number of pages
- iii, 49 pages
- Resource Type
- Thesis
- Language
- English
- Academic Unit
- College of Medicine; Pharmacology and Physiology; Drexel University
- Other Identifier
- 991021229715604721