Molecular Cloning and Expression of Cro s ۱: an Occupational Allergen from Saffron Pollen (Crocus sativus)

Publish Year: 1391
نوع سند: مقاله ژورنالی
زبان: English
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شناسه ملی سند علمی:

JR_RBMB-1-1_001

تاریخ نمایه سازی: 10 شهریور 1400

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 ۱, 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 ۱ (rCro s ۱) was expressed as a soluble protein in Pichia pastoris and purified to homogeneity by gel filtration. Inhibition of IgE binding to rCro s ۱ by pollen extract was analyzed by ELISA. Results: The allergen Cro s ۱ was identified from saffron pollen extracts and cloned by PCR. Cro s ۱ cDNA defined an acidic polypeptide with homology to pollen proteins from Chenopodium album and Ligastrum vulgaris. The rCro s ۱ was expressed in P. pastoris at ۲۸ mg/l. Saffron pollen extract inhibited the binding of patient serum IgE to rCro s ۱. Conclusion: We identified and cloned the first Crocus sativus pollen allergen. rCro s ۱ cDNA shows a very high homology with Che a ۱, the major allergen of lambchr('۳۹')s-quarter, Chenopodium album, Caryophyllales, pollen (۹۷%). Cro s ۱ is a useful tool for specific diagnosis and structural studies of occupational allergy to saffron.

Authors

Abdol-Reza Varasteh

Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran

Mojtaba Sankian

Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran

Terumi Midoro-Horiuti

INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA.

Mohamad Taghi Shakeri

Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran

Edward G. Brooks

Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas, USA

Randall M. Goldblum

Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas, USA

Martin D. Chapman

INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA.

Anna Pomés

INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA.