Identification of polcalcin as a novel allergen of Amaranthus retroflexus pollen
Allergol. immunopatol
; 47(4): 357-364, jul.-ago. 2019. tab, graf
Article
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| IBECS
| ID: ibc-186507
Biblioteca responsable:
ES1.1
Ubicación: BNCS
ABSTRACT
Introduction: Amaranthus retroflexus (Redroot Pigweed) is one of the main sources of allergenic pollens in temperate areas. Polcalcin is a well-known panallergen involved in cross-reactivity between different plants. The aim of this study was the molecular cloning and expression of polcalcin, as well as evaluating its IgE-reactivity with A. retroflexus sensitive patients' sera. Methods: Allergenic extract was prepared from A. retroflexus pollen and the IgE-reactivity profile was determined by ELISA and immunoblotting using sera from twenty A. retroflexus sensitive patients. Polcalcin-coding sequence was amplified by conventional PCR method and the product was inserted into pET-21b(+) vector. The recombinant protein was expressed in E. coli BL21 and purified by metal affinity chromatography. The IgE-binding capability of the recombinant protein was analyzed by ELISA and immunoblotting assays, and compared with crude extract. Results: Of 20 skin prick test positive patients, 17 patients were positive in IgE-specific ELISA. Western blotting confirmed that approximately 53% of ELISA positive patients reacted with 10kDa protein in crude extract. The A. retroflexus polcalcin gene, encoding to 80 amino acid residues was cloned and expressed as a soluble protein and designated as Ama r 3. The recombinant polcalcin showed rather identical IgE-reactivity in ELISA and western blotting with 10 kDa protein in crude extract. These results were confirmed by inhibition methods, too. Conclusion: The recombinant form of A. retroflexus polcalcin (Ama r 3) could be easily produced in E. coli in a soluble form and shows rather similar IgE-reactivity with its natural counterpart
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Colección:
06-national
/
ES
Base de datos:
IBECS
Asunto principal:
Polen
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Proteínas de Unión al Calcio
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Pruebas Cutáneas
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Alérgenos
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Rinitis Alérgica Estacional
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Amaranthus
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Antígenos de Plantas
Límite:
Adolescent
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Adult
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Female
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Humans
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Male
Idioma:
En
Revista:
Allergol. immunopatol
Año:
2019
Tipo del documento:
Article