Studio dell'espressione di una Flavon Diiron Protein (FDP) inGiardia intestinalis

Giardia lamblia is a flagellated unicellular eukaryotic microorganism that commonly causes diarrheal disease throughout the world. AlthoughGiardiahas a poor tolerance to O2, it preferentially colonizes the fairly aerobic upper part of the small intestine.The flavodiiron proteins (FDP) are widespread among strict or facultative anaerobic prokaryotes, where they are involved in the response to nitrosative and/or oxidative stress. The FDP fromGiardiahas high O2-reductase activity but very low NO-reductase activity; O2reacts with the reduced protein quite rapidly and with high affinity producing H2O.Experiments were designed to examine whether the FDP recombinant, stimulate the immune system of the host (mouse) producing specific antibodies. To verify if they specifically recognize the native protein inGiardia, the antibodies have been used to set up by blotting and fluorescence techniques.Experiments were performed under conditions of stress in order to obtain information on the location and expression of the native protein. I wanted to study the overexpression of the protein to determine its intracellular location and confirm the data obtained with not transformed cells.Further characterization experiments have led to study the protein expression during encystations and to evaluating the expression of FDP in metronidazoleresistant cells compared to drug sensitive protozoa.