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Cloning and Characterization of the Amino Terminal 30% of Retinoid and Fatty Acid Binding Glycoprotein in Schneider’s S2 Cells

UNCP Author/Contributor (non-UNCP co-authors, if there are any, appear on document)
Ashley D. Bowman (Creator)
Institution
The University of North Carolina at Pembroke (UNCP )
Web Site: http://www.uncp.edu/academics/library
Advisor
Jeremy Sellers

Abstract: Our lab is interested in exploring insect lipid transporting genes and the lipoproteins they produce within Drosophila melanogaster. We want to study lipid transporting genes in insects because several labs have demonstrated that components necessary for the synthesis and secretion of insect lipoproteins are similar to those required for production of low-density lipoproteins (LDL) in humans. This is of particular importance given that elevated LDL concentration in human plasma is known to lead to atherosclerosis and heart disease. It is our lab’s intent to develop models for the exploration of lipoprotein biogenesis using D. melanogaster in expectation that our findings may contribute to further understanding of human LDL production. At present, our interest centers on the production of insect apolipophorin II/I, named retinoid and fatty acid binding glycoprotein (RFABG) in D. melanogaster. RFABG is a homolog of the major structural protein on human LDLs, apolipoprotein B, and is known to transport fatty acids to insect flight muscle during extended flights. As a result we have chosen to subclone an N-terminal portion of RFABG, which constitutes about 30% of the entire DNA sequence (RF30). Through sequential digests and purification procedures, our lab successfully transferred the entire RF30 sequence into the insect expression vector, pRmHa-3. We have also appended a short, engineered sequence to RF30 that encodes the FLAG epitope tag and a termination codon. Future efforts may now explore expression of this FLAG-tagged RF30 construct in Schneider’s S2 cells to determine whether these cells are capable of producing lipoproteins.

Additional Information

Publication
Honors Project
Language: English
Date: 2009
Keywords
Lipoproteins, Insect Lipoproteins, Low Density Lipoproteins, Atherosclerosis, Drosophia Melanogaster, Retinoid Fatty Acid Glycoprotein