Construction of a Microbacteriophage Lysogen

Abstract

Bacteriophages are viruses that infect and replicate in a bacterial host cell. The Oklahoma soil due to its diversity is a good source for finding new bacteriophages. Due to the emergence of drug-resistant bacteria, there is an increasing need for isolating new phages that can be used for phage therapy. In my research, I have purified and sequenced three bacteriophages from Oklahoma soil. These bacteriophages were previously isolated by Virology students at the University of Central Oklahoma. The soil phages were isolated using Microbacterium foliorum as the host bacteria. I hypothesize that we will be able to find a lysogen for one of the three phages. A lysogen is a host bacterium cell that contains a stably integrated copy of a phage genome. This research began with the purification of each phage sample, followed by amplification to obtain purified phage lysates. Viral genomic DNA was extracted from each of the three samples. Phage genomes were characterized using restriction digest with HaeIII, NspI, and SacII. Phages Arroyo and Busephilis have been sequenced using Illumina sequencing technology. Phages Arroyo and Busephilis are newly discovered bacteriophages from Oklahoma soil with a genome length of 42129 and 52986 bp respectively. I have successfully created a lysogen for Phage Sasian. The purified lysogen can be used to test the ability of other phages to infect the new lysogen. The results from this study expand the knowledge of host immunity to phage infection.