Adapter-Directed Display: A Modular Design for Shuttling Display on Phage Surfaces

A novel adapter-directed phage display system was developed with modular features. In this system, the target protein is expressed as a fusion protein consisting of adapter GR1 from the phagemid vector, while the recombinant phage coat protein is expressed as a fusion protein consisting of adapter GR2 in the helper phage vector. Surface display of the target protein is accomplished through specific heterodimerization of GR1 and GR2 adapters, followed by incorporation of the heterodimers into phage particles. A series of engineered helper phages were constructed to facilitate both display valency and formats, based on various phage coat proteins. As the target protein is independent of a specific phage coat protein, this modular system allows the target protein to be displayed on any given phage coat protein and allows various display formats from the same vector without the need for reengineering. Here, we demonstrate the shuttling display of a single-chain Fv antibody on phage surfaces between multivalent and monovalent formats, as well as the shuttling display of an antigen-binding fragment molecule on phage coat proteins pIII, pVII, and pVIII using the same phagemid vectors combined with different helper phage vectors. This adapter-directed display concept has been applied to eukaryotic yeast surface display and to a novel cross-species display that can shuttle between prokaryotic phage and eukaryotic yeast systems.     

Multiple-population versus hierarchical conifer breeding programs: a comparison of genetic diversity levels

Advanced-generation domestication programs for forest-tree species has raised some concerns about the maintenance of genetic diversity in forest-tree breeding programs. Genetic diversity in natural stands was compared with two genetic conservation options for a third-generation elite Pinus taeda breeding population. The breeding population was subdivided either on the basis of geographic origin and selection goals (multiple-population or MPBS option) or stratified according to genetic value (hierarchical or HOPE option). Most allelic diversity in the natural stands of loblolly pine is present in the domesticated breeding populations. This was true at the aggregate level for both multiple-population (MPBS) and the hierarchical (HOPE) populations. Individual subpopulations within each option had less genetic diversity but it did not decline as generations of improvement increased. Genetic differentiation within the subdivided breeding populations ranged from 1 to 5%, genetic variability is within each subpopulation rather than among subpopulations for both MPBS (>95%) and the HOPE approaches (>98%). Nei's G_st estimates for amongpopulation differentiation were biased upwards relative to estimates of from Weir and Cockerham (1984).