Molecular and Immunochemical ‎Characterization of Pop n 2: A New Allergen ‎of Populus nigra Pollen

Shams, Mohammad‐Hossein and Kheirandish, Farnaz and Varzi ‎, Ali‐Mohammad and Sohrabi ‎, Seyyed‐Mohsen and Safari, Mojgan (2020) Molecular and Immunochemical ‎Characterization of Pop n 2: A New Allergen ‎of Populus nigra Pollen. Clin Exp Allergy.

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Background Pollen is one of the most common allergens that cause respiratory allergies worldwide. Pollen grains from poplars ‎have been reported as important sources of pollinosis in many countries.‎ Objective The aim of the present study was to determine the molecular and immunochemical characterization of Pop n 2, a ‎novel allergen of Populus nigra (P. nigra) pollen extract.‎ Methods In this study, the pollen extract of P. nigra was analyzed by SDS‐PAGE, and the allergenic profile was determined ‎by IgE immunoblotting and specific ELISA using the sera of twenty allergic patients. The coding sequence of Pop n ‎‎2 was cloned and expressed in the Escherichia coli BL21 (DE3) using plasmid the pET‐21b (+). Finally, the ‎expressed recombinant Pop n 2 was purified by affinity‐chromatography.‎ Results Pop n 2 belongs to the profilin family with a molecular weight of approximately 14 kDa. Pop n 2 is the most IgE‐‎reactive protein (about 65%) in the P. nigra pollen extract. The cDNA sequencing results indicated an open reading ‎frame 396 bp that encodes 131 amino acid residues. The results of ELISA and Immunoblotting assays showed that ‎recombinant Pop n 2 could react with the IgE antibody in patients' sera, like its natural counterpart.‎ Conclusion Our data revealed that Pop n 2 is a significant allergen in the P. nigra pollen extract. Moreover, we observed that ‎the recombinant Pop n 2 produced by the pET‐21b (+) vector in the E. coli system acts as its natural counterpart.‎ ‎ ‎

Item Type: Article
Subjects: R Medicine > RB Pathology
Divisions: Faculty of Medicine, Health and Life Sciences > School of Medicine
Depositing User: samira sepahvandy
Date Deposited: 16 Dec 2020 14:44
Last Modified: 16 Dec 2020 14:44

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