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Date of Award

2003

Document Type

Thesis - Pacific Access Restricted

Degree Name

Master of Science (M.S.)

Department

Biological Sciences

First Advisor

Gregg Jongeward

First Committee Member

Craig A. Vierra

Second Committee Member

Lisa Wrischnik

Abstract

The basic helix-loop-helix (bHbH) family-of transcription factors is important in many developmental and regulatory pathways such as cellular proliferation and differentiation, lineage commitment, sex determination, neurogenesis, myogenesis, hematopoeisis and pancreatic development. The free-living nematode Caenorhabdits elegans is an important model organism. Genetics studies of a gene in nematodes help us to better understand the functioning of hornologs in more complex organisms. These studies investigate the nematode homolog of ABF-1, CeABF-1, and its potential role in the development of C. elegans. The BLAST Database (http://www.ncbi.nlm.nih.gov) predicted a bHLH protein in C. elegans, located on cosmid ZK682.4, of 170 amino acids with an overall 51% similarity to human ABF-1. Importantly, it had a 72% similarity to human ABF-1 within the bHLH domain. Genomic and eDNA clones of CeABF-1 were isolated using whole nematodes and PCR methods. Clones were constructed that would allow us to use green fluorescent protein (GFP) to localize CeABF-1 expression. We also used RNA interference to determine the function of CeABF-1. No obvious phenotype was observed in nematodes unable to produce the CeABF-1 protein. Preliminary studies suggest that CeABF-1 is not an essential gene for development in C. elegans. Constructs were also made for protein induction and antibody studies. Further studies are necessary to determine how CeABF-1 is involved in nematode development and its interaction with other proteins.

Pages

86

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