Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/17560
Title: Identifying protein co-factors of oct4, an essential 'stemness' transcription factor, by affinity purification and mass spectrometry
Authors: CHEONG YUJING CLARA
Keywords: ES cell pluripotency, Oct4, Protein Interaction, Epitope Tagging, Affinity Purification, Mass Spectrometry
Issue Date: 28-Jan-2009
Source: CHEONG YUJING CLARA (2009-01-28). Identifying protein co-factors of oct4, an essential 'stemness' transcription factor, by affinity purification and mass spectrometry. ScholarBank@NUS Repository.
Abstract: The demands of embryonic development require tight transcriptional regulation in order to manage multiple signals and outcomes in the organism. Sequence-specific transcription factors play central roles in this coordinative procedure, acting as a molecular switchboard for the control of expression. Oct4 is a key factor governing the pluripotency of embryonic stem cells, which are feted for their capacity to both self-renew and differentiate to all cell lineages of the embryo proper. To gain perspective on the role of Oct4 through its protein interactions, I have engineered ES cell lines expressing epitope b tagged Oct4 from the endogenous locus at biologically relevant levels. Putative partners were isolated by affinity purification and identified by mass spectrometry. In this work, known partners of Oct4 were reaffirmed, and new interactions that pointed towards a spectrum of roles for Oct4 in transcriptional regulation were established.This work addresses two needs b one is the realization of protein interactions mediated by a transcription factor central to the maintenance of pluripotency, and the other is the development of a generic platform for the discovery of protein-protein interactions in a biologically relevant context.
URI: http://scholarbank.nus.edu.sg/handle/10635/17560
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