Construction of a Francisella tularensis 2-DE protein database

Lenka Hernychova, Jiri Stulik, Petr Halada, Ales Macela, Michal Kroca, Torsten Johansson, Michal Malina

Institute of Radiobiology and Immunology, Purkyne Military Medical Academy, Hradec, Kralove, Czech Republic, jstulik@pmfhk.cz

We have started the construction of a two dimensional database of the proteome of Francisella tularensis, a bacterium that is responsible for the highly pathogenic disease tularemia. The genome of this intracellular pathogen is not completely sequenced, yet and, currently, information only about 54 proteins is available from NCBI database. We have analyzed the Francisella tularensis live vaccine strain by two-dimensional gel electrophoresis with immobilized pH 3-10 gradient in the first dimension and 9-16 % gradient or tricine SDS-PAGE in the second dimension. In both cases about 2000 spots were detected. Furthermore, we compared the protein pattern of the non-virulent Francisella tularensis live vaccine strain with protein profiles of two wild type clinical isolates and more than 50 differentially expressed proteins were counted. The separated proteins are going to be identified by peptide mass fingerprinting, however, due to the lack of complete genome sequence data only eight proteins were unambiguously identified, so far. Among them, acid phosphatase and the most basic isoform of a hypothetical 23 kDa protein are characteristic only for virulent strains.