Authors : Nazanin Pazouki , Mojtaba Sankian , Taher Nejadsattari , Ramezan-Ali Khavari-Nejad and Abdol-Reza Varasteh
Abstract: Oriental Plane trees, as Platanus orientalis, are an important source of airborne allergens in cities of the southwest Asia and southeast Europe. Diagnosis of type I allergy essentially depends on the availability of defined allergens, which can be provided by recombinant technology. This study was aimed to investigate molecular properties of a Platanus orientalis pollen allergen, designated as Pla or 1.0101 (accession number EU296476), to produce its immunoreactive recombinant counterpart in Escherichia coli. Molecular characterization of the Platanus orientalis pollen allergen was performed using, cDNA cloning, expression of the recombinant allergen in Escherichia coli and IgE immunoblotting of recombinant allergen. The 18 kDa allergen (Pla or 1.0101) was identified as an important IgE-binding component of Platanus orientalis pollen. The Pla or 1.0101-specific cDNA sequences were amplified, using specific primers based on the N and C-terminal sequence of a Pla or 1.0101 homologue in Platanus acerifolia, Pla a 1. Sequencing corresponding Pla or 1.0101 cDNA revealed an open reading frame of 513 bp coding for 170 amino acid residues. The recombinant Pla or 1.0101 was produced by pET102/D-TOPO E. coli expression system. IgE-binding to the recombinant form of Pla or 1.0101 was proven by immunoblot and specific ELISA.
Nazanin Pazouki , Mojtaba Sankian , Taher Nejadsattari , Ramezan-Ali Khavari-Nejad and Abdol-Reza Varasteh , 2008. Complementary DNA Cloning and Immunologic Characterization of a New Platanus orientalis Pollen Allergen, Pla or 1.0101. Research Journal of Biological Sciences, 3: 1419-1425.