Publication Date

2015

Document Type

Thesis

Committee Members

Madhavi Kadakia (Advisor), Michael Markey (Committee Member), Yong-Jie Xu (Committee Member)

Degree Name

Master of Science (MS)

Abstract

δ NP63α, the primary p63 isoform of the p53 transcription factor family, is a proto-oncogene implicated in non-melanoma skin cancers. Expressed in the basal layer of the epidermis, δ NP63α promotes cell survival and proliferation. Inhibition of this protein could potentially be beneficial in non-melanoma skin cancer patients. The first goal of this project was the purification of recombinant δ NP63α in Escherichia Coli. Recombinant δ NP63α was expressed as GST-δ NP63α followed by GST cleavage using GST trap affinity column chromatography yielding pure δ NP63α The second objective of this project was to test the binding capabilities of peptides previously identified by phage display to δ NP63α. Six 12-mer peptides were previously identified by phage display using filamentous phage M13 followed by biopanning and amplification of target bound peptides. The amplified pools were sequenced and studied. In this study, the identified peptides were linked to biotin, the affinity and specificity of these peptides was evaluated using two different methods, streptavidin Dynabeads and ELISA. Through these methods, the biotinylated peptide P5 was found to be the most specific peptide to bind δ NP63 α . Future results utilizing these techniques may reveal promising therapeutic agents for the treatment of non-melanoma skin cancer.

Page Count

83

Department or Program

Department of Biochemistry and Molecular Biology


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