Streamlined phage display library protocols for identification of insect gut binding peptides highlight peptide specificity

• DOI: 10.1016/j.cris.2021.100012
• Publication Date: 2021-02-21T22:39:24Z
• Authors: Ruchir Mishra; Ya Guo; Pavan Kumar; Pablo Emiliano Ca...; Clebson S. Tavares; Rahul Banerjee; Suyog Kuwar; Bryony C. Bonning
• Keywords: Biopanning; Exigua

Phage display libraries have been used to isolate insect gut binding peptides for use as pathogen transmission blocking agents, and provide artificial anchors increased toxicity of bacteria-derived pesticidal proteins. Previously, phage clones displaying enriched were sequenced by Sanger sequencing. Here we present a streamlined protocol identification peptides, using insect-appropriate feeding strategies, with next generation sequencing tailored bioinformatics analyses. The pipeline is designed eliminate poorly false positive identify predicted be stable hydrophilic. In addition developing protocols, also sought address whether candidate can bind insects from more than one order, which an important consideration safe, practical peptide-modified To this end, screened that the epithelia two pest insects, Asian citrus psyllid, Diaphorina citri (Hemiptera) beet armyworm, Spodoptera exigua (Lepidoptera), beneficial insect, western honey bee, Apis mellifera (Hymenoptera). While unique peptide sequences totaling 13,427 D. citri, 89,561 S. 69,053 A. identified eluted surface guts, final pools comprised 53, 107 1423 respectively. benefits multiple rounds biopanning, along properties in relation proteins control are discussed.