PhiXing-it, displaying foreign peptides on bacteriophage ΦX174

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• 作者：Kristofer J. Christakos^b ; Janice A. Chapman^e ; Bentley A. Fane^a ; ^d ; ^e ; ^{bfane@email.arizona.edu" class="auth_mail" title="E-mail the corresponding author} ; Samuel K. Campos^a ; ^b ; ^c ; ^e ; ^{skcampos@email.arizona.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Microvirus ; ΦX174 ; Phage display ; Peptide display ; Antigen display ; Papillomavirus ; HPV ; HPV16 ; L2 ; RG-1
• 刊名：Virology
• 出版年：2016
• 出版时间：15 January 2016
• 年：2016
• 卷：488
• 期：Complete
• 页码：242-248
• 全文大小：2458 K

文摘

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.