Mycobacteriophage (phage) are a group of viruses that infect bacteria in the genus Mycobacterium. Two phage lifestyles are lytic and temperate. Lytic phage only carry out the lytic life cycle, resulting in host cell lysis. Temperate phage are capable of completing both lytic and lysogenic life cycles. During the lysogenic life cycle, a phage-encoded integrase facilitates integration at sites attP in the phage genome and attB in the host to form a lysogen. The cluster E mycobacteriophage integration system is poorly understood. Ukulele, a lysogenic cluster E phage, is being used to identify the Cluster E attP and characterize lysogeny regulation. A putative attP containing sequence was identified in the Ukulele genome by computational analysis. To confirm the presence of attP, this sequence will be inserted into a plasmid and transferred into integrase expressing M. smegmatis (pST-KT-int). Cells will be screened for plasmid integrated into the genome. To characterize the role of the integrase in lysogeny regulation, we will determine the impact of integrase expression levels on induction event frequency in M. smegmatis (pST-KT-int) – Ukulele lysogens.
Whitaker, Emily E., "Characterization of the Mycobacteriophage Ukulele Integration System; Identification of Integration Site Attp and the Role of the Integrase in Lysogeny Regulation" (2016). Honors College. 414.