Generation of an FtsW-EMGFP recombinant fusion protein in F. tularensis LVS

Francisella tularensis is a tier 1 select agent that causes the potentially fatal disease Tularemia. F. tularensis has been designated a tier 1 agent based on its potential for use as a bioweapon. The CDC describes symptoms of Tularemia that vary from skin ulcers to pneumonia, symptoms differ based on route of infection. F. tularensis is an airborne pathogen, however contact with contaminated water and arthropod bites also risk infection. Relatively little is known about the cell division processes of F. tularensis, however, its genome contains homologs of the key division and cell wall (dcw) genes that have been extensively studied in Escherichia coli. The product of one such gene, FtsW is involved in the recruitment of the transpeptidase FtsI and stabilization of the cytokinetic ring in E. coli. Ftl_1613 is the gene that encodes the putative FtsW homolog in F. tularensis LVS. To learn more about the function of FtsW in F. tularensis LVS, we have amplified Ftl_1613 with specific primers to allow its cloning into the EMGFP reporter plasmids, pSC13 and pSC18. Fluorescence microscopy will be used to analyze the subcellular localization of the resulting FtsW-EMGFP fusion protein and provide insight into the cell division process of F. tularensis. (This work was supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence).