Honors Theses and Capstones

Date Completed

Spring 2025

Abstract

Toxoplasma gondii is an obligate intracellular parasite with a complex life cycle and an unusual transcriptional machinery compared to other eukaryotes. Unlike most organisms, Toxoplasma lacks multiple well-conserved promoter motifs and has multiple TATA-binding proteins (TBPs), including TBP1, whose function in DNA binding and transcription regulation is poorly understood. This study aimed to generate and express a recombinant TBP1 protein from Toxoplasma to enable future DNA-binding assays and motif identification. Using RT-PCR, the TgTBP1 gene was amplified and cloned into a pGEX vector to produce a GST-tagged protein. The recombinant plasmid was transformed into E. coli BL21 cells, and protein expression was induced using IPTG. Expression peaked at 8 and 16 hours post-induction, producing a 57 kDa protein consistent with GST-TBP1. The successful expression of TBP1 lays the groundwork for further studies to elucidate its role in gene regulation in Toxoplasma, including electrophoretic mobility shift assays.

First Advisor

Vicki Jeffers

College or School

COLSA

Department or Program

Biomedical Sciences: Medical and Veterinary Science

Degree Name

Bachelor of Science

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