title: Molecular cloning and expression of Cro s 1: an occupational allergen from saffron pollen (Crocus sativus).
abstract: 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.
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran.
authors.forename: Abdol-Reza
authors.initials: AR
authors.lastname: Varasteh
authors.affiliation: Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Sankian
authors.affiliation: INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA.
authors.forename: Terumi
authors.initials: T
authors.lastname: Midoro-Horiuti
authors.affiliation: Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran.
authors.forename: Malihe
authors.initials: M
authors.lastname: Moghadam
authors.affiliation: Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran.
authors.forename: Mohamad Taghi
authors.initials: MT
authors.lastname: Shakeri
authors.affiliation: Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas, USA.
authors.forename: Edward G
authors.initials: EG
authors.lastname: Brooks
authors.affiliation: Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas, USA.
authors.forename: Randall M
authors.initials: RM
authors.lastname: Goldblum
authors.affiliation: INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA.
authors.forename: Martin D
authors.initials: MD
authors.lastname: Chapman
authors.affiliation: INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA.
authors.forename: Anna
authors.initials: A
authors.lastname: Pomés
keywords: Allergen; Cro s 1; Occupational allergy; Saffron pollen; cDNA cloning