title: Frameshifting in the p6 cDNA phage display system.
abstract: Phage display is a powerful technique that enables easy identification of targets for any type of ligand. Targets are displayed at the phage surface as a fusion protein to one of the phage coat proteins. By means of a repeated process of affinity selection on a ligand, specific enrichment of displayed targets will occur. In our studies using C-terminal display of cDNA fragments to phage coat protein p6, we noticed the occasional enrichment of targets that do not contain an open reading frame. This event has previously been described in other phage display studies using N-terminal display of targets to phage coat proteins and was due to uncommon translational events like frameshifting. The aim of this study was to examine if C-terminal display of targets to p6 is also subjected to frameshifting. To this end, an enriched target not containing an open reading frame was selected and an E-tag was coupled at the C-terminus in order to measure target display at the surface of the phage. The tagged construct was subsequently expressed in 3 different reading frames and display of both target and E-tag measured to detect the occurrence of frameshifting. As a result, we were able to demonstrate display of the target both in the 0 and in the +1 reading frame indicating that frameshifting can also take place when C-terminal fusion to minor coat protein p6 is applied.
authors.affiliation: Biomedical Research Institute, School of Life Sciences, Transnationale Universiteit Limburg and Hasselt University, Diepenbeek, Belgium. veerle.somers@uhasselt.be.
authors.forename: Cindy
authors.initials: C
authors.lastname: Govarts
authors.forename: Klaartje
authors.initials: K
authors.lastname: Somers
authors.forename: Piet
authors.initials: P
authors.lastname: Stinissen
authors.forename: Veerle
authors.initials: V
authors.lastname: Somers
mesh_terms: D017104:Bacteriophage M13 / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000821:virology; D016368:Frameshift Mutation; D015723:Gene Library; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics*; D019151:Peptide Library; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: C440769:M13 phage protein VI; D019151:Peptide Library; D014760:Viral Fusion Proteins