Slide 1

Slide 2 Unknown Phage 11 Leah Liston Slide 3 Cluster Identification Unknown 11 is identified with Cluster Q Other Phage in Cluster Q Long time singleton: Giles Evanesce HH92 OBUPride Slide 4 Plaque Observations Small and round Slightly turbid Cluster Q Lightly turbid plaques Small Slide 5 Phamerator - Phams Pham 1278 is unique to Cluster Q Evanesce Gene 11 (441bp) Giles Gene 10 (441bp) HH92 Gene 10 (441bp) OBUPride Gene 10 (441bp) The amino acid sequence: (K)AAGAIADASTDEAAAAK(A) At beginning of the genome located after the terminases, protease, portal, and capsid genes Slide 6 Pham 1278 In most phages, between the capsid protein and the tail assembly chaperones proteins there is usually major tail subunit proteins No gene in Giles can be predicted Pham 1278 protein associated with the unpredicted major tail subunit gene Unique gene because of this Unknown 11 from Mass Spec had 97% alignment Biggest Cluster Q indicator Slide 7 Spectrum Mill Compared against phage database top scores Evanesce gene 11 Pham 1278 Cluster Q specific Evanesce gene 10 Pham 1326 Cluster Q specific Evanesce gene 25 Pham 3145 Cluster Q specific Evanesce gene 16 Pham 1283 Cluster Q specific Evanesce gene 40 Pham 1292 Cluster Q specific Bxz2 gene 23 Pham 1842 Not in Cluster Q Evanesce gene 9 Pham 1325 Cluster Q specific Evanesce gene 7 Pham 1311 Cluster P & Q only DUF935 Unknown function but consists of several bacterial protein including Bacteriophage Mu gp29 Bxz2 gene 17 Pham 1841 Not in Cluster Q Slide 8 Spectrum Mill Compared against Mycobacterium Database Evanesce gene 11 Pham 1278 Cluster Q specific Evanesce gene 10 - Pham 1326 Cluster Q specific Elongation factor Tu M. trivale RNA polymerase beta M. chitae Endopeptidase Clp 1 Mycle Glyceraldehyde-2-phosphate dehydrogenase M. tuberculosis H37Rv Evanesce gene 25 Pham 3145 Cluster Q specific ATP-dependent Clp protease M. tuberculosis CDC1551 Alcohol dehydrogenase M. smegmatis Slide 9 Spectrum Mill Results Phage Database High alignments with Evanesce genes High alignment with Pham 1278 Cluster Q specific High alignment with Cluster Q specific genes Mycobacterium Database Alignment with elongation factor Critical function: in translation energy source for binding of tRNA to the A site in ribosomes Endopeptidase Clp 1 Major role in degradation of misfolded proteins Slide 10 Pipeline Comparison with Positive Control Czyszczon1 Both Contain Elongation Factor Tu Chaperonin GroEL Glyceraldehyde-3-phosphate dehydrogenase Fatty acid synthetase I Glutamine synthetase GLNA1 Evanesce Gene 22 Pham 3143 Only in Cluster Q on Phamerator possible mutations Only Unknown 11 Glycerol kinase RNA polymerase beta Evanesce Genes: 17, 43, 11, 10, 21 Slide 11 Chromatography Graphs Unknown 11 Positive Control Czyszcon1 Slide 12 Future Direction Phamerator Look at cluster Q genes that are shared in other phages Gene Function? Slide 13 References Hatfull, G. The Secret Lives of Mycobacteriophages. (2012). Advances in Virus Research Hatfull, G. Comparative genomic analysis of sixty mycobacteriophage genomes: genome clustering, gene acquisition and gene size. (2010). J Mol Biol, 397(1), 119-143 Spectrum Mill Phamerator NCBI Blastp alignments