title: PhiXing-it, displaying foreign peptides on bacteriophage ΦX174.
abstract: Although bacteriophage φX174 is easy to propagate and genetically tractable, it is use as a peptide display platform has not been explored. One region within the φX174 major spike protein G tolerated 13 of 16 assayed insertions, ranging from 10 to 75 amino acids. The recombinant proteins were functional and incorporated into infectious virions. In the folded protein, the peptides would be icosahedrally displayed within loops that extend from the protein׳s β-barrel core. The well-honed genetics of φX174 allowed permissive insertions to be quickly identified by the cellular phenotypes associated with cloned gene expression. The cloned genes were easily transferred from plasmids to phage genomes via recombination rescue. Direct ELISA validated several recombinant virions for epitope display. Some insertions conferred a temperature-sensitive (ts) protein folding defect, which was suppressed by global suppressors in protein G, located too far away from the insertion to directly alter peptide display.
authors.affiliation: The Department of Molecular and Cellular Biology, The University of Arizona, Tucson, AZ, USA.
authors.forename: Kristofer J
authors.initials: KJ
authors.lastname: Christakos
authors.affiliation: The BIO5 Institute, The University of Arizona, Tucson, AZ, USA.
authors.forename: Janice A
authors.initials: JA
authors.lastname: Chapman
authors.affiliation: The Department of Immunobiology, The University of Arizona, Tucson, AZ, USA; The School of Plant Sciences, The University of Arizona, Tucson, AZ, USA; The BIO5 Institute, The University of Arizona, Tucson, AZ, USA. Electronic address: bfane@email.arizona.edu.
authors.forename: Bentley A
authors.initials: BA
authors.lastname: Fane
authors.affiliation: The Department of Immunobiology, The University of Arizona, Tucson, AZ, USA; The Department of Molecular and Cellular Biology, The University of Arizona, Tucson, AZ, USA; The Cancer Biology Graduate Interdisciplinary Program, The University of Arizona, Tucson, AZ, USA; The BIO5 Institute, The University of Arizona, Tucson, AZ, USA. Electronic address: skcampos@email.arizona.edu.
authors.forename: Samuel K
authors.initials: SK
authors.lastname: Campos
mesh_terms: D010584:Bacteriophage phi X 174 / Q000235:genetics*; D061505:Cell Surface Display Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D016254:Mutagenesis, Insertional; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics*; D011995:Recombination, Genetic; D014764:Viral Proteins / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins
keywords: Antigen display; HPV; HPV16; L2; Microvirus; Papillomavirus; Peptide display; Phage display; RG-1; ΦX174