Characterization of the low-level expression of HIV-1 HXB2CG wild-type envelope glycoprotein gp41 ectodomain in an Escherichia coli expression system
Characterization of the low level expression of HIV-1 HXB2CG wild-type envelope glycoprotein gp41 ectodomain in an Escherichia coli expression system
M.S. (Master of Science)
Department of Chemistry and Biochemistry
HIV (Viruses); Viral envelopes; Glycoproteins; Escherichia coli
This thesis concerns the HIV - 1 viral envelope glycoprotein gp41 which is responsible for one of the initial stages of HIVinfection, the fusion of the viral envelope and the host cell membrane. Initially a structural dynamic study of the protein was to be performed, but the focus of the project shifted as the previously unstudied full-length wild-type gp41 ectodomain was not able to be synthesized in the E. coli strains used to grow recombinant protein. Four cloning vectors were generated, and none were found to be stable enough in the cell line BL21 DE3 pLysS to generate quantities of protein necessary for further experimental work. Plasmid instability is evidenced by instability plating tests, the resurgence of mutant nonexpressing plasmids postinduction, and the low total yield of protein: lOOng/ gram of wet cells. Reasons for instability may include the presence of rare codons in the target protein and potential oligomerization of the amphipathic helical protein with host proteins. Further experimental work on this protein requires overcoming the problem of plasmid instability.
Nezrick, Tania A., "Characterization of the low-level expression of HIV-1 HXB2CG wild-type envelope glycoprotein gp41 ectodomain in an Escherichia coli expression system" (2000). Graduate Research Theses & Dissertations. 1751.
ix, 79 pages
Northern Illinois University
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Includes bibliographical references (pages -59)