Research Interests Mycobacterium tuberculosis kills more people than any other single infectious agent. Since antibiotics are available and the BCG vaccine is in widespread use, why do nearly three million people die each year from TB? The answer, in part, is that we really don't understand this curious bacterium or what parts of its genetic instructions make this such a deadly pathogen. At the heart of our strategies to understand mycobacterial genetics is the mycobacteriophages - viruses that infect the mycobacteria. These are easy to grow and manipulate and offer advantages over working with the slow-growing mycobacteria (such as M. tuberculosis) that can take up to a month to produce a colony on an agar plate. Phages are also rich sources of a wide variety of potential genetic and molecular tools that can be used to study - and to modify - their bacterial hosts. Here's just a flavor of some of the current studies going on in the lab : Bacteriophage genomics. We have sequenced about half-a-dozen mycobacteriophage genomes and, in collaboration with Dr. Hendrix, an equivalent number of other phage genomes. These studies have not only provided some valuable insights into how this incredible collection of organisms have evolved, but fuels our mycobacterial investigations by continually providing new phage systems to study. Mycobacterial Gene Expression. We have identified novel strategies that the mycobacteria use for regulating gene expression. We have used these for developing tools for controlled expression of genes in mycobacteria and are now delving into the mechanisms of how these system works. Site-specific recombination. Several of the mycobacteriophages that we are investigating integrate their DNA into the host chromosome (and can excise them too). We are building a variety of mycobacterial vectors that use these systems to permit the integration of any desirable genes into mycobacterial genomes. We are also using these as model systems to investigate the molecular mechanisms involved in site-specific recombination and attempting to elucidate how phages dictate the directionality of these events. Tools - Genetic and Clinical. Studying the mycobacteria and their phages has great potential for the development of novel tools for their genetics but also for a more direct clinical involvement. Two systems we have been involved in developing are multivalent recombinant BCG vaccines and Luciferase Reporter Phages, but there are numerous additional strategies awaiting further development!   Selected Publications Ojha AK, Baughn AD, Sambandan D, Hsu T, Trivelli X, Guerardel Y, Alahari A, Kremer L, Jacobs WR Jr, Hatfull GF. Growth of Mycobacterium tuberculosis biofilms containing free mycolic acids and harbouring drug-tolerant bacteria. Mol Microbiol. 2008 Jul;69(1):164-74. van Kessel JC, Hatfull GF. Mycobacterial recombineering. Methods Mol Biol. 2008;435:203-15. Ghosh P, Bibb LA, Hatfull GF. Two-step site selection for serine-integrase-mediated excision: DNA-directed integrase conformation and central dinucleotide proofreading. Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3238-43. Epub 2008 Feb 25. van Kessel JC, Hatfull GF. Efficient point mutagenesis in mycobacteria using single-stranded DNA recombineering: characterization of antimycobacterial drug targets. Mol Microbiol. 2008 Mar;67(5):1094-107. Epub 2008 Jan 22. Morris P, Marinelli LJ, Jacobs-Sera D, Hendrix RW, Hatfull GF. Genomic characterization of myco-bacteriophage Giles: Evidence for phage acquisition of host DNA by illegitimate recombination. J Bacteriol. 2008 Mar;190(6):2172-82. Epub 2008 Jan 4. Complete Publication Listing   Grant Support NIH: Integration and excision by serine-integrases. Principal Investigator NIH: Phage mimicry of microbacterial signaling. Principal Investigator NIH: Mycobacteriophage mediated recombineering in Mycobacterium Tuberculosis. Principal Investigator Howard Hughes Medical Institute: HHMI's Professors Program and HHMI UG Science Education Program. Principal Investigator NSF: Microbial genome sequencing: Sequencing of exceptionally large bacteriophage. Co-Investigator   Other Links Dept. of Biological Sciences University of Pittsburgh

Graham F. Hatfull, Ph.D. Office:  376 Crawford Hall Lab:376 Crawford Hall Phone:(412) 624-6975 Fax: (412) 624-4870 gfh+@pitt.edu   Academic Affiliations Chair, Dept. of Biological Sciences HHMI Professor Eberly Family Professor of Biotechnology   Education 1981 - Ph.D. Molecular Biology University of Edinburg Edinburg, UK 1981-1983 - Postdoc Molecular Biology Yale University 1983 - 1984 Postdoc Molecular Biology MRC (LMB) Cambridge, UK 1984 - 1988 Postdoc Molecular Biology Yale University   Lab Personnel Postdoctoral Fellows: Dr. Gregory Broussard, Dr. Pallavi Ghosh, Dr. Anil Ohja, Dr. Mariana Piuri, Dr. Zuzana Swigonova Graduate Students: Amrita Balachandran, Kim Clemens, Laura Marinelli, Julia van Kessel Undergrad Researchers: Michael Davies, Andrew Hrychowian Staff: Mridula Anand, Deborah Jacobs-Sera Administrator for the Chair of Biological Sciences: Crystal Petrone Phone: 412-624-4350