A new phage display system using bacteriophage phi6

Phage display is a useful molecular technique for a wide range of applications, including protein-protein interactions, in vitro protein evolution (i.e. protein engineering), and drug discovery. However, there are some limitations of the current technology that are in urgent need of improvement. The membrane proteome accounts for 23% of all the proteins encoded by the human genome. This becomes an underexplored area as membrane proteins are intricate in terms of their structural and functional properties. It is therefore necessary to introduce a new phage display system specific for membrane proteins. 

This study employed the bacteriophage φ6 of the family Cystoviridae, a group of enveloped viruses that have a tripartite dsRNA genome and a unique lipid bilayer derived from the host plasma membrane. By adapting and modifying an existing reverse-genetics system for the rescue of bacteriophage φ6, I have demonstrated that the host Pseudomonas syringae strain HB10Y was able to maintain a plasmid construct harbouring a fusion gene of phage protein P9 to a membrane protein HTR3A followed by a six-histidine tag. Furthermore, bacteriophage φ6 was able to conduct replication in the presence of the plasmid. These preliminary results will lay a foundation for further development of a novel phage display system.