Honors Theses and Capstones

Date of Award

Spring 2025

Project Type

Senior Honors Thesis

College or School

COLSA

Department

Molecular, Cellular, and Biomedical Sciences

Program or Major

Biomedical Sciences: Medical and Veterinary Science

Degree Name

Bachelor of Science

First Advisor

Vicki Jeffers

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.

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