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Isolation, cloning, and characterization of the 2S albumin: A new allergen from hazelnut by Cristiano Garino; Laurian Zuidmeer; Justin Marsh; Alison Lovegrove; Maria Morati; Serge Versteeg; Piet Schilte; Peter Shewry; Marco Arlorio; Ronald van Ree is a Medicine article available to read on EtoBox.
What is Isolation, cloning, and characterization of the 2S albumin: A new allergen from hazelnut about?
## Abstract **Scope:** 2S albumins are the major allergens involved in severe food allergy to nuts, seeds, and legumes. We aimed to isolate, clone, and express 2S albumin from hazelnut and determine its allergenicity. **Methods:** 2S albumin from hazelnut extract was purified using size exclusion chromatography and RP‐HPLC. After __N__‐terminal sequencing, degenerated and poly‐d(T) primers were used to clone the 2S albumin sequence from hazelnut cDNA. After expression in __Escherichia coli__ and affinity purification, IgE reactivity was evaluated by Immunoblot/ImmunoCAP (inhibition) analyses using sera of nut‐allergic patients. **Results:** __N__‐terminal sequencing of a ∼10 kDa peak from size exclusion chromatography/RP‐HPLC gave two sequences highly homologous to pecan 2S albumin, an 11 amino acid (aa) __N__‐terminal and a 10aa internal peptide. The obtained clone (441 bp) encoded a 147aa hazelnut 2S albumin consisting of a putative signal peptide (22 aa), a linker peptide (20 aa), and the mature protein sequence (105 aa). The latter was successfully expressed in __E. coli__. Both recombinant and natural 2S albumin demonstrated similar IgE reactivity in Immunoblot/ImmunoCAP (inhi
Who reads Isolation, cloning, and characterization of the 2S albumin: A new allergen from hazelnut?
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- Author
- Cristiano Garino; Laurian Zuidmeer; Justin Marsh; Alison Lovegrove; Maria Morati; Serge Versteeg; Piet Schilte; Peter Shewry; Marco Arlorio; Ronald van Ree
- Publisher
- John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley (ISSN 1613-4125)
- Published
- 2010
- Language
- EN
- Field
- Medicine (Health Sciences)