Hertveldt Kirsten, Beliën Tim, Volckaert Guido
Department of Biosystems, Division of Gene Technology, Katholieke Universiteit Leuven, Leuven, Belgium.
Methods Mol Biol. 2009;502:321-39. doi: 10.1007/978-1-60327-565-1_19.
In M13 phage display, proteins and peptides are exposed on one of the surface proteins of filamentous phage particles and become accessible to affinity enrichment against a bait of interest. We describe the construction of fragmented whole genome and gene fragment phage display libraries and interaction selection by panning. This strategy allows the identification and characterization of interacting proteins on a genomic scale by screening the fragmented "proteome" against protein baits. Gene fragment libraries allow a more in depth characterization of the protein-protein interaction site by identification of the protein region involved in the interaction.
在M13噬菌体展示中,蛋白质和肽段暴露于丝状噬菌体颗粒的一种表面蛋白上,从而可针对感兴趣的诱饵进行亲和富集。我们描述了片段化全基因组和基因片段噬菌体展示文库的构建以及通过淘选进行的相互作用筛选。该策略通过针对蛋白质诱饵筛选片段化的“蛋白质组”,从而在基因组规模上鉴定和表征相互作用蛋白。基因片段文库通过鉴定参与相互作用的蛋白质区域,能够更深入地表征蛋白质-蛋白质相互作用位点。