Biochemistry and Molecular Biology-BS
Biochemistry and Molecular Biology
Dr. John Tansey
First Committee Member
Dr. Jennifer Bennett
Second Committee Member
Dr. Margaret Koehler
Perilipin 5, Alternative Splicing, Northern Blot, Diabetes, Gene Expression, Lipid Droplet
Biochemistry | Molecular Biology
Alternative splicing occurs throughout the human genome, leading to multiple proteins from a single gene. The resulting proteins can be nearly identical or vastly different in how they function. Alternative splicing within the perilipin family has been observed in perilipins 1 and 3, giving rise to proteins with varying functions. In the course of our studies on perilipin 5, an immunoblot signal was observed that corresponded to an approximately 35 kDa protein. Western blot analysis of this protein has revealed its expression in C2C12 cultured myoblasts and in heart and liver mouse tissue. Further analysis of perilipin 5 cDNA through reverse transcriptase PCR provided evidence that this protein is a result of alternative splicing occurring in perilipin 5 that we have termed perilipin 5b. The objective of the current study is to develop DNA based probes for use in northern blot analysis of perilipin 5. The genomic DNA sequence of perilipin 5 was analyzed to identify areas where splicing events could occur, and probes were designed accordingly to detect the different variants of perilipin 5. In the analysis of different mouse tissue samples, these probes have been used to detect which of the perilipin 5 splice variants are being expressed under different feeding conditions. We hypothesize that probes made to detect the 5’ end of perilipin 5 will detect both variants, while probes for the 3’ end will selectively bind to one variant.
Ranzau, Brodie, "Evidence of a Perilipin 5 Splice Variant" (2017). Honors Thesis Projects. 55.
Available for download on Saturday, April 13, 2019