Alteration of tail fiber protein gp38 enables T2 phage to infect Escherichia coli O157:H7

• Published in: Journal of biotechnology Vol. 115; no. 1; pp. 101 - 107
• Main Authors: Yoichi, Masatoshi, Abe, Michiharu, Miyanaga, Kazuhiko, Unno, Hajime, Tanji, Yasunori
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 01.01.2005; Amsterdam Elsevier; New York, NY
• Subjects: Bacteriophage; Bacteriophage T4 - pathogenicity; Bacteriophage T4 - physiology; Bacteriophage T4 - ultrastructure; Biological and medical sciences; Biotechnology; Escherichia coli; Escherichia coli - cytology; Escherichia coli - virology; Escherichia coli O157:H7; Fundamental and applied biological sciences. Psychology; LPS; Mutagenesis, Site-Directed; Phage T2; Protein Engineering - methods; Receptor; Recombinant Proteins - metabolism; Tail fiber; Viral Tail Proteins - genetics; Viral Tail Proteins - metabolism; Tail fiber; Bacteriophage; Receptor; Escherichia coli O157:H7; LPS; Escherichia coli; Receiver; Myoviridae; Protein; Virus; Infection; Phage T2; T even phage; Bacteria; Phage; Enterobacteriaceae
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2004.08.003

Artificial control of phage specificity may contribute to practical applications, such as the therapeutic use of phages and the detection of bacteria by their specific phages. To change the specificity of phage infection, gene products (gp) 37 and 38, expressed at the tip of the long tail fiber of T2 phage, were exchanged with those of PP01 phage, an Escherichia coli O157:H7 specific phage. Homologous recombination between the T2 phage genome and a plasmid encoding the region around genes 37– 38 of PP01 occurred in transformant E. coli K12 cells. The recombinant T2 phage, named T2ppD1, carried PP01 gp37 and 38 and infected the heterogeneous host cell E. coli O157:H7 and related species. On the other hand, T2ppD1 could not infect E. coli K12, the original host of T2, or its derivatives. The host range of T2ppD1 was the same as that of PP01. Infection of T2ppD1 produced turbid plaques on a lawn of E. coli O157:H7 cells. The binding affinity of T2ppD1 to E. coli O157:H7 was weaker than that of PP01. The adsorption rate constant ( k a) of T2ppD1 (0.17 × 10 −9 (ml CFU −1 min −1) was almost 1/6 that of PP01 (1.10 × 10 −9 (ml CFU −1 min −1)). In addition to the tip of the long tail fiber, exchange of gene products expressed in the short tail fiber may be necessary for tight binding of recombinant phage.