Rhodobacter Capsulatus Bacteriophage Isolation, DNA Extraction, and DNA Sequencing

Presenter and Advisor Information

Max Garcia, Illinois Wesleyan University

Submission Type

Event

Faculty Advisor

David Bollivar

Expected Graduation Date

2018

Location

Center for Natural Sciences, Illinois Wesleyan University

Start Date

4-21-2018 2:00 PM

End Date

4-21-2018 3:00 PM

Disciplines

Education

Abstract

Ten million trillion trillion bacteriophages inhabit the earth and survive by targeting bacteria. Bacteriophages are viruses that infect bacteria. Although there are large numbers of bacteriophages, only a subset of these will infect a particular species of bacteria. When trying to discover new phages, selecting a host bacterium affects what bacteriophages can be isolated. TheYW1 strain of Rhodobacter capsulatus was chosen as the host for this experiment because this bacterial species has different methods for metabolism and is used as a model organism to study processes such as as bacterial photosynthesis. Water samples were taken from a sewage treatment plant in Bloomington Illinois and mixed with host cells. Areas of phage infection, called plaques, were found. Plaques are formed because once the virus enters the cell it ultimately will cause the death of the bacterial cell. Different strains of the bacterium R.capsulatus were tested to demonstrate if the isolated phages were similar in what hosts they infect. The plaques were also used to extract DNA of the phages to allow for DNA sequencing and characterization of the phage genome. Discovering novel phages is the main focus for this research and was eventually achieved through isolating phages, extracting phage DNA, and conducting host range tests that demonstrate the different strains of Rhodobacter capsulatus that are infected by the phage.

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Apr 21st, 2:00 PM Apr 21st, 3:00 PM

Rhodobacter Capsulatus Bacteriophage Isolation, DNA Extraction, and DNA Sequencing

Center for Natural Sciences, Illinois Wesleyan University

Ten million trillion trillion bacteriophages inhabit the earth and survive by targeting bacteria. Bacteriophages are viruses that infect bacteria. Although there are large numbers of bacteriophages, only a subset of these will infect a particular species of bacteria. When trying to discover new phages, selecting a host bacterium affects what bacteriophages can be isolated. TheYW1 strain of Rhodobacter capsulatus was chosen as the host for this experiment because this bacterial species has different methods for metabolism and is used as a model organism to study processes such as as bacterial photosynthesis. Water samples were taken from a sewage treatment plant in Bloomington Illinois and mixed with host cells. Areas of phage infection, called plaques, were found. Plaques are formed because once the virus enters the cell it ultimately will cause the death of the bacterial cell. Different strains of the bacterium R.capsulatus were tested to demonstrate if the isolated phages were similar in what hosts they infect. The plaques were also used to extract DNA of the phages to allow for DNA sequencing and characterization of the phage genome. Discovering novel phages is the main focus for this research and was eventually achieved through isolating phages, extracting phage DNA, and conducting host range tests that demonstrate the different strains of Rhodobacter capsulatus that are infected by the phage.