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en Molecular Cloning and Expression of Cro s 1: an Occupational Allergen from Saffron Pollen (Crocus sativus) ایمنی شناسی Immunology Background: The cultivation of saffron is expanding through the southeast of Iran, and allergy to saffron pollen occurs in workers involved in processing this plant. We aimed to clone, sequence and express a major allergen involved in saffron pollen allergy, and to compare the recombinant with the natural allergen. Methods: The N-terminal amino acid sequence of Cro s 1, an allergen from saffron pollen, was determined after immunoblotting. The cDNA encoding for this allergen was cloned by PCR utilizing a primer based on the N-terminal amino acid sequence. Recombinant Cro s 1 (rCro s 1) was expressed as a soluble protein in Pichia pastoris and purified to homogeneity by gel filtration. Inhibition of IgE binding to rCro s 1 by pollen extract was analyzed by ELISA. Results: The allergen Cro s 1 was identified from saffron pollen extracts and cloned by PCR. Cro s 1 cDNA defined an acidic polypeptide with homology to pollen proteins from Chenopodium album and Ligastrum vulgaris. The rCro s 1 was expressed in P. pastoris at 28 mg/l. Saffron pollen extract inhibited the binding of patient serum IgE to rCro s 1. Conclusion: We identified and cloned the first Crocus sativus pollen allergen. rCro s 1 cDNA shows a very high homology with Che a 1, the major allergen of lamb's-quarter, Chenopodium album, Caryophyllales, pollen (97%). Cro s 1 is a useful tool for specific diagnosis and structural studies of occupational allergy to saffron. Allergen, cDNA cloning, Cro s 1, Occupational allergy, Saffron pollen 1 8 http://rbmb.net/browse.php?a_code=A-10-1-1&slc_lang=en&sid=1 Abdol-Reza Varasteh varasteha@mums.ac.ir 10031947532846005881 10031947532846005881 Yes Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran Mojtaba Sankian 10031947532846005882 10031947532846005882 No Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran Terumi Midoro-Horiuti 10031947532846005883 10031947532846005883 No INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA. Malihe Moghadam 10031947532846005884 10031947532846005884 No Mohamad Taghi Shakeri 10031947532846005885 10031947532846005885 No Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran Edward G. Brooks 10031947532846005886 10031947532846005886 No Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas, USA Randall M. Goldblum 10031947532846005887 10031947532846005887 No Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas, USA Martin D. Chapman 10031947532846005888 10031947532846005888 No INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA. Anna Pomés 10031947532846005889 10031947532846005889 No INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA.