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Detection of a putative Brugia malayi POU-homeodomain transcription factor binding to its congate promoter
Steven A. Williams
Bachelor of Arts
Neglected tropical diseases, Lymphatic filariasis, Drug target candidate analysis, Brugia malayi, Transcription factor and promoter interaction, Binding analysis
Neglected tropical diseases (NTDs) mainly burden individuals living predominantly in developing countries. One of the most detrimental NTDs is lymphatic filariasis (LF), a parasitic disease that results in the development of elephantiasis and is caused by the parasitic nematodes: Wuchereria bancrofti, Brugia malayi, and Brugia timori. With increasing prevalence of drug resistant human filarial nematodes, the need for novel therapeutics to eradicate LF has become crucial. Despite the urgent necessity to understand gene expression in filarial parasites for identification of new therapeutic targets, little is known about promoters and transcription factors in these organisms. The overall aim of this study is to identify possible transcription factors in B. malayi, using molecular and biochemical techniques, that could serve as potential drug targets to help combat filariasis. A comparative analysis between the protein sequence datasets of B. malayi and Caenorhabditis elegans identified a putative transcription factor Bma-UNC-86 and its cognate promoter mec-3. Electrophoretic mobility shift assay (EMSA) is being used to confirm the specific binding of the UNC-86 protein to the mec-3 promoter DNA and quantitively determine an apparent binding constant. This will allow us to design a DNA affinity chromatography assay that will be used to capture the Bma-UNC-86 transcription factor from total protein extracted from B. malayi and other putative transcription factors that recognize the mec-3 promoter. Conformation of binding between UNC-86 and mec-3 promoter, successful predication of apparent DNA dissociation (KD) constant, and use of a DNA affinity chromatography will provide a pool of potential parasite specific drug targets to combat filarial parasites.
©2019 Hafsa Mohamud Mire. Access limited to the Smith College community and other researchers while on campus. Smith College community members also may access from off-campus using a Smith College log-in. Other off-campus researchers may request a copy through Interlibrary Loan for personal use.
Mire, Hafsa Mohamud, "Detection of a putative Brugia malayi POU-homeodomain transcription factor binding to its congate promoter" (2019). Honors Project, Smith College, Northampton, MA.
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118 pages : illustrations (some color), Includes bibliographical references (pages 114-118)