Research Publication Title

The Isolation, Purification and DNA Analysis of a Potentially Novel Mycobacteriophage

Major

Biology

Faculty Mentor

Indiren Pillay

Keywords

bacteriophage, DNA, virus, mycobacterium, restriction digest

Abstract

Bacteriophages are viruses that infect and lyse bacteria with a high degree of specificity. These ubiquitous viruses not only help to maintain natural balances of microbial populations but have also been quintessential in refining our understanding of horizontal gene transfer as well as microbial evolution. Mycobacteriophages are viruses that infect the bacterial genus, Mycobacterium, which include species like Mycobacterium tuberculosis and Mycobacterium smegmatis. The role of mycobacteriophages in controlling certain mycobacterial infections is of great interest. As a result of bacteriophage’s ability to proliferate and evolve rapidly, phages maintain significant genetic diversity. These genetic variations create characteristic interactions between the virus and its host cell. In order to examine how phages interact with the non-pathogenic host bacteria, M. smegmatis, mycobacteriophages were isolated and purified from a sedimentary sample. The sample was collected from soil in the Georgia College Teaching Greenhouse. Soil samples were enriched and mycobacteriophages were subsequently isolated and purified. Purified phage demonstrated a lytic infection cycle of its host. The phage DNA was subsequently extracted, subjected to restriction enzyme digestion, and analyzed through gel electrophoresis. Comparative analysis could help to uncover functions associated with distinctive proteins in the isolated phage’s DNA. Understanding these protein functions has further implications in determining efficacy of a phage on the removal of biofilms associated with infections.

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The Isolation, Purification and DNA Analysis of a Potentially Novel Mycobacteriophage

Bacteriophages are viruses that infect and lyse bacteria with a high degree of specificity. These ubiquitous viruses not only help to maintain natural balances of microbial populations but have also been quintessential in refining our understanding of horizontal gene transfer as well as microbial evolution. Mycobacteriophages are viruses that infect the bacterial genus, Mycobacterium, which include species like Mycobacterium tuberculosis and Mycobacterium smegmatis. The role of mycobacteriophages in controlling certain mycobacterial infections is of great interest. As a result of bacteriophage’s ability to proliferate and evolve rapidly, phages maintain significant genetic diversity. These genetic variations create characteristic interactions between the virus and its host cell. In order to examine how phages interact with the non-pathogenic host bacteria, M. smegmatis, mycobacteriophages were isolated and purified from a sedimentary sample. The sample was collected from soil in the Georgia College Teaching Greenhouse. Soil samples were enriched and mycobacteriophages were subsequently isolated and purified. Purified phage demonstrated a lytic infection cycle of its host. The phage DNA was subsequently extracted, subjected to restriction enzyme digestion, and analyzed through gel electrophoresis. Comparative analysis could help to uncover functions associated with distinctive proteins in the isolated phage’s DNA. Understanding these protein functions has further implications in determining efficacy of a phage on the removal of biofilms associated with infections.

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