Developments in the use of baculoviruses for the surface display of complex eukaryotic proteins

The ability to couple genotype to phenotype has proven to be of immense value in systems such as phage display and has allowed genes encoding novel functions to be selected directly from complex libraries. However, the complexity of many eukaryotic proteins places a severe constraint on successful display in Escherichia coli. This restriction could be resolved if a eukaryotic virus could be similarly engineered for display purposes. Preliminary data have suggested that the baculovirus Autographa californica, a multiple nuclear polyhedrosis virus (AcMNPV) is a candidate for eukaryotic virus display because the insertion of peptides into the native virus coat protein, or the expression of foreign proteins as coat protein fusions, results in incorporation of the sequence of interest onto the surface of virus particles. A variety of strategies are currently under investigation to develop further the display capabilities of AcMNPV and to improve the complexity of library that might be accommodated. Several expression vectors for different forms of surface display have been developed and, coupled with improved recombination strategies, represent progress towards a refined tool for use in functional genomics and in vitro protein evolution.