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.

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Apr 26th, 11:05 AM Apr 26th, 11:20 AM

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.