MunkgeeRoachy: A Gordonia rubripertincta Bacteriophage Isolated from Soil
Abstract
There are many bacteria that are growing resistant to antibiotics. Therefore, there is an urgent need for alternative therapies. Bacteriophage therapy, the use of viruses to treat bacterial infections, is a promising alternative. The goal of this project was to isolate and identify phages from soil samples that infect Gordonia rubripertincta. G. rubripertincta is a gram-positive bacterium that is typically found in soil and aquatic habitats. Gordonia species are rapidly becoming associated with human and animal diseases. Infections of G. rubripertincta have typically been found after various surgical procedures such as the applications of catheters, or the use of heart-lung machines. In this experiment, a soil sample was collected at Wolf Lake Park in Davie, Florida to isolate bacteriophage that eliminate G. rubripertincta. The phage was isolated from the soil sample using a direct isolation method. Phage purification was conducted to ensure that unique phage was present in the sample. To purify the phage, isolated plaques were picked, diluted, and subjected to additional plaque assays. The phage (MungkeeRoachy) was successfully isolated after three rounds of purification. To produce a high concentration of phage, plaque assays were repeated, and the phage lysate was collected. Once the hightiter lysate was obtained, the phage was characterized by its genome. The DNA was extracted and then sent for sequencing. The phage sample of MunkgeeRoachy was imaged with Cryo-EM. Based on morphology, they were classified as being siphoviridae. Future applications can entail using the phages found as an advantage for bacteriophage therapy.
Faculty Sponsors
Dr. Julie Torruellas Garcia
Project Type
Event
Location
Alvin Sherman Library
Start Date
4-5-2023 12:00 PM
End Date
4-6-2023 4:00 PM
MunkgeeRoachy: A Gordonia rubripertincta Bacteriophage Isolated from Soil
Alvin Sherman Library
There are many bacteria that are growing resistant to antibiotics. Therefore, there is an urgent need for alternative therapies. Bacteriophage therapy, the use of viruses to treat bacterial infections, is a promising alternative. The goal of this project was to isolate and identify phages from soil samples that infect Gordonia rubripertincta. G. rubripertincta is a gram-positive bacterium that is typically found in soil and aquatic habitats. Gordonia species are rapidly becoming associated with human and animal diseases. Infections of G. rubripertincta have typically been found after various surgical procedures such as the applications of catheters, or the use of heart-lung machines. In this experiment, a soil sample was collected at Wolf Lake Park in Davie, Florida to isolate bacteriophage that eliminate G. rubripertincta. The phage was isolated from the soil sample using a direct isolation method. Phage purification was conducted to ensure that unique phage was present in the sample. To purify the phage, isolated plaques were picked, diluted, and subjected to additional plaque assays. The phage (MungkeeRoachy) was successfully isolated after three rounds of purification. To produce a high concentration of phage, plaque assays were repeated, and the phage lysate was collected. Once the hightiter lysate was obtained, the phage was characterized by its genome. The DNA was extracted and then sent for sequencing. The phage sample of MunkgeeRoachy was imaged with Cryo-EM. Based on morphology, they were classified as being siphoviridae. Future applications can entail using the phages found as an advantage for bacteriophage therapy.
