Date of Award
Master of Arts
Dr. Gyula Ficsor
Dr. Karim Essani
Dr. W. Gary Tarpley
Masters Thesis-Open Access
The human immunodeficiency virus (HIV) is the etiological agent of the acquired immune deficiency syndrome (AIDS). This retrovirus, encodes a RNA dependent DNA polymerase, reverse transcriptase (RT), which is essential to virus replication. Biochemical analysis indicated that RT is composed of 2 polypeptides of 66,000 and 51,000 daltons (p66 and p51) which combine to form a heterodimer. This heterodimer is thought to arise from the post-translational processing where p66 is made and either autocatalytic or proteolytic cleavage events result in a heterogeneous p66/p51. Site-directed mutagenesis was used at the proposed processing site to probe HIV-RT processing.
Three prokaryotic expression vectors each carrying a mutated form of RT were constructed. These mutants were induced and the product evaluated for RT enzymatic activity and the forms of RT protein visualized on SDS polyacrylamide gels and western blots. Results suggest that the mutations were at or near the RT processing site.
Poppe, Susan M., "Site-Directed Mutagenesis of the Putative Processing Site of Recombinant Human Immunodeficiency Virus Reverse Transcriptase" (1989). Master's Theses. 4537.