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Frameshifting in the P6 cDNA Phage Display System
Biomedical Research Institute, School of Life Sciences, Transnationale Universiteit Limburg and Hasselt University, Diepenbeek, Belgium
* Author to whom correspondence should be addressed.
Received: 9 November 2010; in revised form: 3 December 2010 / Accepted: 16 December 2010 / Published: 20 December 2010
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.
Keywords: cDNA phage display; E. coli; frameshifting; p6; filamentous phage
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Cite This Article
MDPI and ACS Style
Govarts, C.; Somers, K.; Stinissen, P.; Somers, V. Frameshifting in the P6 cDNA Phage Display System. Molecules 2010, 15, 9380-9390.
Govarts C, Somers K, Stinissen P, Somers V. Frameshifting in the P6 cDNA Phage Display System. Molecules. 2010; 15(12):9380-9390.
Govarts, Cindy; Somers, Klaartje; Stinissen, Piet; Somers, Veerle. 2010. "Frameshifting in the P6 cDNA Phage Display System." Molecules 15, no. 12: 9380-9390.