Control of pathogen by bacteriophage carrying variable host range constructed by molecular evolution engineering

2016 Fiscal Year Final Research Report

• Project/Area Number: 24246133
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: Tanji Yasunori 東京工業大学, 生命理工学院, 教授 (00282848)
• Co-Investigator(Kenkyū-buntansha): 宮永 一彦 東京工業大学, 生命理工学研究科, 助教 (40323810)
• Research Collaborator: Hidetomo Iwano 酪農学園大学, 教授
• Project Period (FY): 2012-04-01 – 2017-03-31
• Keywords: バクテリオファージ / ファージセラピー / 黄色ブドウ球菌 / 薬剤耐性菌 / 乳房炎 / 大腸菌

Outline of Final Research Achievements

The emergence of antibiotic-resistant bacteria has increased the attention to the therapeutic use of bacteriophages. In order to improve the usability of phages, it is required to understand the mechanism underlying host recognition, especially receptor-binding proteins (RBPs) which determine the host range. In this study, we demonstrate staphylococcal phages ΦSA012 possesses at least two RBPs. A polyclonal antibody against ORF103 inhibited infection of ΦSA012 in the presence of α-GlcNAc, suggesting that ORF103 (ligand) binds to α-GlcNAc (Receptor).
Phage-resistant E. coli isolated from co-culture with phage T2 was used. As the target of modification, Hyper Variable Region (HVR) was selected. This region is part of gene38, which encodes the tip of the long tail fiber. CRISPR/Cas system facilitated homologous recombination between wild type phage and plasmid which encoded random mutation in the HVR. Artificially modified phage showed infectivity to the phage resistant E.coli.

Free Research Field

生物化学工学