Thesis
Expression and purification of the hepatitis B virus X protein
Master of Science (M.S.), Drexel University
Aug 2017
DOI:
https://doi.org/10.17918/etd-7748
Abstract
The hepatitis B virus (HBV) genome is made of a small partially double-stranded, circular DNA that encodes four open-reading frames. One important protein encoded by the smallest open reading frame of the viral genome is HBV protein X; this protein has been shown to be essential for HBV replication. It has also been shown that the X protein plays an important role in the development of HBV-associated HCC. To this date, there is no three-dimensional structure available for HBx, and this may be due to the presence of disordered regions in HBx. Understanding the structure of X is essential for understanding its ordered and disordered boundaries as well as when using in-vitro assays to understand HBx functions. We propose expressing different truncated version of protein X in bacterial system, using different vectors that help stabilize the protein during purification. We expressed eight different versions of protein X plus, including the full-length protein, in three different vectors using BL21 bacterial cells as hosts. Our goal was to test the expression of these versions of the protein using small test-expression to see if any of the regions of the protein can be expressed. We use the same conditions used on large-scale expression. Our results show low expression of three different constructs in one of the vectors used in this experiment. Conditions can be optimized to yield higher expression. Overall, we have promising results that if optimized could lead to a better expression and for the first time give us information about HBx three-dimensional structure.
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Details
- Title
- Expression and purification of the hepatitis B virus X protein
- Creators
- Guila Fakhoury - DU
- Contributors
- Patrick J. Loll (Advisor) - Drexel University (1970-)
- Awarding Institution
- Drexel University
- Degree Awarded
- Master of Science (M.S.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Number of pages
- 73 pages
- Resource Type
- Thesis
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems (1997-2026); Drexel University
- Other Identifier
- 7748; 991014632292904721