Bacteriophages (viruses of bacteria) vary with respect to the collection of strains of bacteria they can attack, a character termed host range. A generalist is a bacteriophage that can attack and successfully kill multiple bacteria, perhaps across even several genera, whereas a specialist is a bacteriophage that can successfully kill a smaller number of bacterial strains. The genetic basis of these differences is not well understood. The isolation of a collection of wild bacteriophage that are able to attack at least one strain of Bacillus subtilis will be done. Then we will assess the ability of these phages to attack additional B. subtilis strains. By sequencing the genomes of phage with different abilities to attack few, or many, bacterial strains, we will be able to search for putative genes, which correlate with these differences. Gene prediction will employ bioinformatics techniques that analyze genomic DNA sequences for features characteristic of open reading frames and hence putative genes. By examining the similarity between these putative genes and sequences stored in the National Center of Biotechnology Information’s database, it may be possible to identify the functions of these ecologically important sequences. In detail, bacteriophage will be isolated by incubating soil with bacterial growth media and then filtering the culture with a filter size that will remove bacteria but allow phage to pass. Single phage from this filtrate will be isolated via dilution of the filtrate and plating on bacterial lawns at dilutions that yield individual phage plaques. Phage from these single plaques with be collected and phage numbers amplified via additional bacterial infection yielding sufficient material for host range assays and genomic sequencing.

Analyzing correlations between genetics and ecology of Big Bend bacteriophages

2019 Spring National Meeting

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