Isolation and Characterization of Gordonia Bacteriophages Laura JoAn Powell, Umar Sahi, Hari Kotturi, and Ralph E. Jones University of Central Oklahoma, Edmond, OK 73034

Abstract

Bacteriophages (phages) are viruses, known to infect bacterial hosts. Many bacteria are used in a variety of areas such as nutrition, farming, and bioremediation. Our host bacteria is Gordonia terrae, found in soils, which has been investigated for bioremediation use. The purpose of this research is isolation and characterization of phages specific to Gordonia. We hypothesize that we should be able to isolate unique Gordonia phages from various soil samples. Gordonia terrae was grown on PYCa media at 28° C. Soil samples were processed using direct and enriched isolation. A plaque assay was performed to detect plaques (voids) in the host bacterial lawn, indicating bacterial cell death. Plaques from each sample were purified and amplified. Purified phages were viewed with a Transmission Electron Microscope to determine the phage morphology. The PCI-SDS method was used for DNA extraction, prior to sequencing. Restriction analysis was done using restriction enzymes. Our preliminary results suggest that we were able to isolate and purify two Gordonia bacteriophages from two soil samples. Both bacteriophages have Siphoviridae morphology with a long and flexible tail. The DNA of isolated phages will be sequenced to determine the uniqueness of our phages.