title: Development of a plasmid display system using GAL4 DNA binding domain for the in vitro screening of functional proteins.
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.
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, 151-742, Seoul, Korea.
authors.forename: Yoo Seong
authors.initials: YS
authors.lastname: Choi
authors.forename: Seung Pil
authors.initials: SP
authors.lastname: Pack
authors.forename: Young Je
authors.initials: YJ
authors.lastname: Yoo
mesh_terms: D003001:Cloning, Molecular; D019151:Peptide Library; D010957:Plasmids / Q000235:genetics*; D011506:Proteins / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics
chemical_list: D019151:Peptide Library; D011506:Proteins; D011993:Recombinant Fusion Proteins