Articles
USING TWO-DIMENSIONAL GEL ELECTROPHORESIS APPROACH FOR CHARACTERIZING OF THE OLE E 1, AN OLIVE POLLEN MAJOR ALLERGEN
Article number
932_9
Pages
69 – 72
Language
English
Abstract
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.
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.
Authors
A. Soleimani, J.D. Alché, A.J. Castro, M.I. Rodríguez-García, A.R. Ladan Moghadam
Keywords
Olea europaea L.
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