Development of a Plasmid Display System using GAL4 DNA Binding
Domain for the <Emphasis Type="Italic">in Vitro</Emphasis> Screening of Functional Proteins |
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Authors: | Yoo?Seong?Choi Seung?Pil?Pack Email author" target="_blank">Young?Je?YooEmail author |
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Institution: | (1) School of Chemical and Biological Engineering, Seoul National University, 151-742 Seoul, Korea;(2) International Innovation Center, Kyoto University, 611-0011 Kyoto, Japan |
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Abstract: | A plasmid display system using GAL4 DNA binding domain (GAL4 DBD) was constructed to enrich the molecular diversity and in vitro selection of functional proteins. Model proteins used were enhanced green fluorescent protein (EGFP) and glutathione S-transferase (GST). The feasibility of this display system was examined using enrichment experiments of target protein from
a model protein mixture and identifying the encoding genes by PCR, in which the model protein mixture includes GAL4 DBD/GST
fusion protein, GAL4 DBD/EGFP fusion protein, and xylanase. Target proteins of GAL4 DBD/GST and GAL4 DBD/EGFP from the model
protein mixture were efficiently isolated by the plasmid display, respectively. The results show that the display system is
sufficiently sensitive to select a target protein from a protein mixture, and that it is possible to discover the functional
proteins from large libraries using relatively simple approaches. |
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Keywords: | GAL4 DNA binding domain in vitro selection plasmid display screening |
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