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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/11513

Title: Frameshifting in the P6 cDNA Phage Display System
Authors: GOVARTS, Cindy
SOMERS, Klaartje
STINISSEN, Piet
SOMERS, Veerle
Issue Date: 2010
Publisher: MDPI
Citation: MOLECULES, 15(12). p. 9380-9390
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.
URI: http://hdl.handle.net/1942/11513
DOI: 10.3390/molecules15129380
ISI #: 000285709000056
ISSN: 1420-3049
Category: A1
Type: Journal Contribution
Validation: ecoom, 2012
Appears in Collections: Research publications

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