Genome and Proteome of Campylobacter jejuni Bacteriophage NCTC 12673

Abstract

Campylobacter jejuni continues to be the leading cause of bacterial food-borne illness worldwide, so improvements to current methods used for bacterial detection and disease prevention are needed. We describe here the genome and proteome of C. jejuni bacteriophage NCTC 12673 and the exploitation of its receptor-binding protein for specific bacterial detection. Remarkably, the 135-kb Myoviridae genome of NCTC 12673 differs greatly from any other proteobacterial phage genome described (including C. jejuni phages CP220 and CPt10) and instead shows closest homology to the cyanobacterial T4-related myophages. The phage genome contains 172 putative open reading frames, including 12 homing endonucleases, no visible means of packaging, and a putative trans-splicing intein. The phage DNA appears to be strongly associated with a protein that interfered with PCR amplification and estimation of the phage genome mass by pulsed-field gel electrophoresis. Identification and analyses of the receptor-binding protein (Gp48) revealed features common to the Salmonella enterica P22 phage tailspike protein, including the ability to specifically recognize a host organism. Bacteriophage receptor-binding proteins may offer promising alternatives for use in pathogen detection platforms.Campylobacter belongs to the epsilon class of proteobacteria and is the leading cause of bacterial food-borne gastroenteritis worldwide (22). Campylobacter has also been associated with severe neurological disorders such as the Guillain-Barré and Miller-Fisher syndromes, as well as reactive arthritis and irritable bowel syndrome (32). Many Campylobacter phages have been characterized by their host-range characteristics, morphology, genome size, and susceptibility to restriction endonucleases (5,7,24). Only a few of the characterized Campylobacter phages are members of the B1 group of the family Siphoviridae (1), while most belong to the family Myoviridae possessing genomes which fall into three size classes: 110 to 150 kb (class III), 170 to 190 kb (class II), and 320 kb (class I) (24). One interesting characteristic of many of these phages is that their DNA appears to be resistant to cleavage by several common restriction endonucleases, thus making them "refractory to genomic analysis" (34). Two genomes of Campylobacter phages have been sequenced thus far: CPt10 and CP220 (34). Both appear to be closely related to each other and belong to the class II of Campylobacter Myoviridae phage genomes (34).Bacteriophages possess enormous diagnostic and therapeutic potential, which provides promise for the development of a wide range of novel antimicrobials and diagnostic tools (14). Numerous phages have been isolated against Campylobacter for use in phage typing schemes (9,10,12,18,25), but it has only recently been shown that bacteriophages can effectively decrease Campylobacter contamination (4, 20, 36). For example, Campylobacter-specific phages were able to reduce the numbers of Campylobacter coli and Campylobacter jejuni in chickens when added directly to the...

Campylobacter jejuni Motility Is Required for Infection of the Flagellotropic Bacteriophage F341

“…4E), as also observed by others (for an example, see reference 33). Moreover, nonadsorbed phages with contracted tail tips and deformed empty heads adsorbed to spherical vesicles were also seen as previously reported for C. jejuni phages (32,33,67) (Fig. 4E).…”

Quo vadis? — MonitoringCampylobacterin Germany

“…[31]). Hence, current research aims to understand the molecular and phenotypic variety of different types of natural bacteriophages [32][33][34][35] in order to rationally design an appropriate cocktail for efficient reduction of Campylobacter in practice.…”

Phages as Therapeutic Tools to Control Major Foodborne Pathogens: Campylobacter and Salmonella

“…The DNA of most Campylobacter phages is difficult to extract, clone and sequence and is refractory to restriction enzyme digestion, which is probably due to tightly adherent and proteinase K resistant proteins Hammerl et al, 2011;Kropinski et al, 2011;Timms et al, 2010). As a consequence, the genome sequence of only five Campylobacter phages have been reported so far Hammerl et al, 2011;Kropinski et al, 2011;Timms et al, 2010).…”

“…As a consequence, the genome sequence of only five Campylobacter phages have been reported so far Hammerl et al, 2011;Kropinski et al, 2011;Timms et al, 2010). Interestingly, the phage genomes known are all related and also part of the T4 superfamily of phages (Petrov et al, 2010).…”