A novel helper phage for Halo Tag-mediated co-display of enzyme and substrate on phage

作者：Delespaul Wouter; Peeters Yves; Herdewijn Piet; Robben Johan^*

来源：Biochemical and Biophysical Research Communications, 2015, 460(2): 245-249.

DOI：10.1016/j.bbrc.2015.03.019

• Phage display
• Molecular evolution
• HaloTag
• Polymerase
• Co-display
• Self-tagging

Phage display is an established technique for the molecular evolution of peptides and proteins. For the selection of enzymes based on catalytic activity however, simultaneous coupling of an enzyme and its substrate to the phage surface is required. To facilitate this process of co-display, we developed a new helper phage displaying HaloTag, a modified haloalkane dehalogenase that binds specifically and covalently to functionalized haloalkane ligands. The display of functional HaloTag was demonstrated by capture on streptavidin-coated magnetic beads, after coupling a biotinylated haloalkane ligand, or after on-phage extension of a DNA oligonucleotide primer with a biotinylated nucleotide by phi29 DNA polymerase. We also achieved co-display of HaloTag and phi29 DNA polymerase, thereby opening perspectives for the molecular evolution of this enzyme (and others) towards new substrate specificities.

• 出版日期2015-5-1
• 单位河北医科大学