USING TWO-DIMENSIONAL GEL ELECTROPHORESIS APPROACH FOR CHARACTERIZING OF THE OLE E 1, AN OLIVE POLLEN MAJOR ALLERGEN

Ole e 1 is a member of the “pollen proteins of the Ole e 1 family” which produces IgE-reactivity in >70% of patients allergic to olive pollen. In order to precisely determine the nature of the differences in the protein expression pattern of Ole e 1 for cultivars of different characteristics, ‘Rowghani’ and ‘Zard’, crude protein was extracted, and isoelectric focusing (IEF) was performed in a IPGphor device (Pharmacia), and analytical gel electrophoresis was also performed according to Laemmli. Gels were transferred onto PVDF membrane and then probed with a monoclonal antibody to Ole e 1. An Alexa Fluor 488 anti-mouse IgG antibody made in goat (molecular probes) was used at as the secondary antibody. Approximately 750 spots could be detected by silver staining in the 2-D map of each sample. Up to 9 of these spots were immunoreactive to the Ole e 1 antibody after Western blotting experi¬ments in ‘Rowghani’, whereas up to five immunoreactive spots only were detected in ‘Zard’. The high degree of polymorphism displayed by Ole e 1 in a cultivar-specific manner may represent part of a unique recognition system. Moreover, its probable relation to different biological behaviors of corresponding cultivars is discussed.