Isolation, identification, and in silico analysis of mycobacteriophage OKCentral2016.

Abstract

Bacteriophages or phages are specific viruses that are capable of infecting bacterial cells without harming eukaryotic cells. Bacteriophages have been utilized to treat and eliminate bacterial infections, which is known as phage therapy. Phages are the most diverse and abundant group of organisms. Mycobacteriophages are a specific type of phage that infect bacteria belonging to the genus Mycobacterium. Mycobacteriophages have been studied due to their potential in killing virulent mycobacteria, such as the causative agents of tuberculosis and leprosy. Mycobacterium smegmatis (M. smegmatis) mc2155 has been used as the model organism for studying mycobacteria due to its increased growth rate and non-pathogenicity. M. smegmatis mc2155 is commonly used to isolate mycobacteriophages from soil. Comparative genomic studies of mycobacteriophages have revealed vast diversity among their genomes. The mycobacteriophage OKCentral2016 was isolated from enriched soil obtained at the University of Central Oklahoma. OKCentral2016 produced transparent plaques, and had a morphology consistent with the Siphoviridae morphotype. The phage remained stable at temperatures below 55??C and under neutral pH conditions. The replication cycle took approximately 4 hours to complete under ideal growing conditions. I also examined the effect of OKCentral2016 on biofilms. OKCentral2016 did not infect any pathogenic mycobacteria tested (M. avium, M. abscessus, M. intracellulare, M. kansasii, and M. simiae). However, it did decrease biofilm formation by the host bacteria. Genomic analysis showed that OKCentral2016 closely resembles phages belonging to the subcluster A10. The genome of OKCentral2016 has a GC content of 65.1%, which codes for 83 open reading frames (ORFs). The annotated genomic sequence is available on GenBank (MF773750) and has been published (1). OKCentral2016 is notable because it was the first mycobacteriophage to be isolated, sequenced, and annotated from Oklahoma soil.