Publication Date
1-1-2000
Document Type
Dissertation/Thesis
Degree Name
B.S. (Bachelor of Science)
Legacy Department
Department of Biological Sciences
Abstract
The Vif (viral infectivity factor) protein of HIV-1 enhances viral infectivity during virus production 100 to 1000 fold (4). Though Vif is required for HIV-1 infection in peripheral blood T lymphocytes, macrophages, and some T cell lines, its exact role during HIV-1 infection remains unknown (13). Currently, it is proposed that Vif may affect the late phases of the viral life cycle, such as virion assembly and maturation (4). In order to get a better understanding of the function and mechanism of Vif, obtaining the structure of the protein is of extreme importance. Unfortunately, no structural information is available. Determining the structure of a protein using NMR (nuclear magnetic resonance) spectroscopy requires that the protein be monomeric in solution. However, Vif exists in solution as aggregates, making NMR impossible. Constructing deletion mutants of Vif may change the protein’s aggregation behavior to monomeric in solution. Thus, these mutants may be the key to determining the structure of Vif, which, in turn, may lead to a better understanding of its function and indicate possible HIV-1 drug targets. PCR (polymerase chain reaction) was used to make the Vif deletion mutants. Restriction digests were performed on both the vector (pDlOVif) and the PCR product using the enzymes BamHl and Hindlll. A ligation and transformation were performed, and transformants were selected for using Ampicillin resistance. A plasmid prep was performed and another restriction digest was performed and run on an agarose gel to ensure the presence of the insert. The Vif mutant was then sequenced as an extra precaution before performing a protein expression test to determine whether or not the protein was soluble.
Recommended Citation
Wehrheim, Courtney, "Deletion mutants may be the key to determining the structure of the Vif protein of HIV" (2000). Honors Capstones. 382.
https://huskiecommons.lib.niu.edu/studentengagement-honorscapstones/382
Extent
12 unnumbered pages
Language
eng
Publisher
Northern Illinois University
Rights Statement
In Copyright
Rights Statement 2
NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.
Media Type
Text
