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Consumption of water contaminated with pathogenic bacteria is a major cause of water-borne diseases. To address this challenge, we have developed a novel and sensitive sensing scheme for the rapid detection of bacteria (Escherichia coli B40) on a fiber-optic platform using bacteriophage (T4) as a bio-recognition element. The novelty of our sensing scheme is that instead of bacteriophages, bacteria (analyte) were first captured on the sensing surface and then the sensing surface was subjected to bacteriophages for specific detection of bacteria. The sensor was subjected to 100 to 10 7 cfu/mL of E. coli B40 spiked in a lake water matrix, and the least concentration of bacteria that could be easily detected was found to be 1000 cfu/mL. The control studies were performed with nonhost bacteria Pseudomonas aeruginosa. Bacteriophage T4, being specific to its host E. coli B40, did not interact with P. aeruginosa captured on the sensing probe, giving a negligible nonspecific response. Due to the specificity of bacteriophages to its host bacteria, it is possible to use this scheme to carry out the detection of specific bacteria in a mixed sample (containing a combination of bacteria) using bacteriophages specific to it. The sensor was able to detect E. coli B40 (target bacteria) even in the presence of a very high concentration (1000 times higher) of P. aeruginosa (nontarget bacteria).

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Genome packaging is a critical step in the assembly of dsDNA bacteriophages and is carried out by a powerful molecular motor known as the large terminase. To date, wild-type structures of only two large terminase proteins are available, and more structural information is needed to understand the genome-packaging mechanism. Towards this goal, the large and small terminase proteins from bacteriophage N4, which infects the Escherichia coli K12 strain, have been cloned, expressed and purified. The purified putative large terminase protein hydrolyzes ATP, and this is enhanced in the presence of the small terminase. The large terminase protein was crystallized using the sitting-drop vapour-diffusion method and the crystal diffracted to 2.8 Å resolution using a home X-ray source. Analysis of the X-ray diffraction data showed that the crystal belonged to space group P222, with unit-cell parameters a = 53.7, b = 93.6, c = 124.9 Å, α = β = γ = 90°. The crystal had a solvent content of 50.2% and contained one molecule in the asymmetric unit.

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