1. A Genomic Comparison of Cluster A Mycobacteriophages: Redefining Homoimmunity Amanda Scott D. Lovas, M. Richters, S. Bhuiyan, B. Miller, J Harmson C.R. Gissendanner, A.M.D. Wiedemeier, A.M. Findley

2. What is a Mycobacteriophage? Virus that infects bacteria, specifically mycobacteria Mycobacteria is a genus of gram negative, acid fast, Actinobacteria Includes M. tuberculosis, M. leprae Also M. smegmatis (fast growing, non-pathogenic)

3. Lytic Bacteriophage Growth

4. Phage Therapy Treatment of Disease with bacteriophage Developed in the 1920s, mainly in the Soviet Union High therapeutic index, few side effects Phage chosen would have to infect target bacteria but be harmless to the organism and normal flora

5. Focusing on A Cluster We worked with sequencing phage that infect M. smeg and may infect M. tuberulosis Compared genome length and genes present Worked with homoimmunity Made a phylogenetic tree that showed degree of divergence

6. Plaque Picture and EM Lysogen Immunity Assays of Mycobacteriophage Eagle, A4 iCleared produces lytic plaques. Isolated following enrichment protocol from a an oak tree litter sample in Rayville, LA. Medusa stained with uranyl acetate and viewed with TEM. Taken at LSU Vet School Scale bar = 0.5μm

7. Restriction Digest Restriction digest of Medusa. Digest comparison of Peaches (left) and Trike (right).

8. Blast

9. Functional Gene Calls 35% of genes in Medusa have identifiable functions

10. Phamerator Comparison of A10 phage Trike (top) only has 63 genes and is missing the repressor binding site

11. Homoimmunity 13bp TGTCAAG is invariable

12. Splitstree for A Cluster phage A1 A7 A4 A10 A3 A2 A6 A9 A5 A8

13. Acknowledgments ULM HHMI Phage Hunters Program 2011- 2012, 2012-2013 This work was supported by the HHMI – SEA Phage Hunters and Louisiana INBRE Programs. We wish to thank Dr. Russell Minton for SPLITSTREE analysis of Cluster A phages.

14. Questions?