Cloning, Expression, & Purification of the Plasmodium Falciparum hypoxanthine guanine xanthine phosphoribosyltransferase to use in Drug Binding Studies
Advisor(s)
Angela Ahrendt, PhD., Illinois Mathematics and Science Academy
Location
Room B133
Start Date
26-4-2019 11:05 AM
End Date
26-4-2019 11:20 AM
Abstract
In this project, the aim is to find a drug to treat malaria. The target that was chosen is an enzyme of Plasmodium falciparum, hypoxanthine guanine xanthine phosphoribosyltransferase (HGXPRT). HGXPRT is an enzyme required for P. falciparum to synthesize purine nucleoside monophosphates essential for DNA/RNA production. When HGXPRT is inhibited there is a potential for P. falciparum to be eliminated. A plasmid containing the gene for HGXPRT was acquired and transformed into BL21 DE3 for expression. The HGXPRT enzyme was expressed and purified using immobilized nickel ion chromatography, and successful expression was confirmed by SDS-PAGE. In addition, the gene was amplified from the plasmid by PCR to insert into a different expression vector to compare the expression between the two systems.
Cloning, Expression, & Purification of the Plasmodium Falciparum hypoxanthine guanine xanthine phosphoribosyltransferase to use in Drug Binding Studies
Room B133
In this project, the aim is to find a drug to treat malaria. The target that was chosen is an enzyme of Plasmodium falciparum, hypoxanthine guanine xanthine phosphoribosyltransferase (HGXPRT). HGXPRT is an enzyme required for P. falciparum to synthesize purine nucleoside monophosphates essential for DNA/RNA production. When HGXPRT is inhibited there is a potential for P. falciparum to be eliminated. A plasmid containing the gene for HGXPRT was acquired and transformed into BL21 DE3 for expression. The HGXPRT enzyme was expressed and purified using immobilized nickel ion chromatography, and successful expression was confirmed by SDS-PAGE. In addition, the gene was amplified from the plasmid by PCR to insert into a different expression vector to compare the expression between the two systems.