Department

Departments of Microbiology and Molecular Biology

First Advisor

Dr. John D. Willford

Description

Ba cteriophage, viruses that specifically infect bacteria, are one of the most prevalent biological entities. Typically, the type of bacteria which a phage is able to infect (host range) is dependent upon the tail fiber which the phage produces. These tail fibers initiate attachment with the bacterial cell and are vital to regular infection. Therefore, the modification or genetic transfer of the tail fiber gene loci allows for modification of the bacteriophage host range. A previous study involved isolation of the tail fiber gene cassette from bacteriophage AR1, and recombining it into bacteriophages T4K10 and T2 in order to modify the T4K10 and T2 host ranges. This study focused on confirming this recombination utilizing phenotypic and genotypic methods. Extensive host range studies where completed using 120 strains of Escherichia coli representing multiple serotypes and 8 negative control species. Each of the recombinant ARX phages created displayed a host range similar to that of AR1 (infecting up to 80 more stains than T4K10 or T2), while maintaining some of the infectivity of the o riginal T4K10 or T2 phages. This hybrid host range was the expected result as has been described previously in the scientific literature. Genetic analyses via restriction fragment length polymorphisms have yet to be successful.

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Poster Presentation, Wyoming I NBRE

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Confirmation of the Gene Cassette Swap of AR1 Tail Fibers to T4K10 Bacteriophage Using Genotypic and Phenotypic Methods

Ba cteriophage, viruses that specifically infect bacteria, are one of the most prevalent biological entities. Typically, the type of bacteria which a phage is able to infect (host range) is dependent upon the tail fiber which the phage produces. These tail fibers initiate attachment with the bacterial cell and are vital to regular infection. Therefore, the modification or genetic transfer of the tail fiber gene loci allows for modification of the bacteriophage host range. A previous study involved isolation of the tail fiber gene cassette from bacteriophage AR1, and recombining it into bacteriophages T4K10 and T2 in order to modify the T4K10 and T2 host ranges. This study focused on confirming this recombination utilizing phenotypic and genotypic methods. Extensive host range studies where completed using 120 strains of Escherichia coli representing multiple serotypes and 8 negative control species. Each of the recombinant ARX phages created displayed a host range similar to that of AR1 (infecting up to 80 more stains than T4K10 or T2), while maintaining some of the infectivity of the o riginal T4K10 or T2 phages. This hybrid host range was the expected result as has been described previously in the scientific literature. Genetic analyses via restriction fragment length polymorphisms have yet to be successful.