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Luud J. W. J. Gilissen, PhD, Suzanne T. H. P

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1 Silencing the major apple allergen Mal d 1 by using the RNA interference approach 
Luud J.W.J. Gilissen, PhD, Suzanne T.H.P. Bolhaar, MD, PhD, Catarina I. Matos, MSc, Gerard J.A. Rouwendal, MSc, Marjan J. Boone, BSc, Frans A. Krens, PhD, Laurian Zuidmeer, PhD, Astrid van Leeuwen, BSc, Jaap Akkerdaas, MSc, Karin Hoffmann-Sommergruber, PhD, André C. Knulst, MD, PhD, Dirk Bosch, PhD, W. Eric van de Weg, PhD, Ronald van Ree, PhD  Journal of Allergy and Clinical Immunology  Volume 115, Issue 2, Pages (February 2005) DOI: /j.jaci Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

2 Fig 1 The construct for gene silencing was built up by linkage of fragment 1 (obtained by PCR using primer 1 [All1HpaI; p1] and primer 2 [All2SmaI; p2] and fragment 2 (from primer 1 [All1HpaI; p1] and primer 3 [All3SmaI; [p3]) at the SmaI site and insertion into the expression cassette pRAP 37. UTR, Untranslated region. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

3 Fig 2 SPT reactivity of 3 transformants and 3 control plantlets in 3 patients. Reactivity was expressed relative to the histamine control. Pt, Patient. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

4 Fig 3 Cross-reactivity of the monoclonal antibody 5H8 (directed to Bet v 1) (A) and of human IgE (B) to Mal d 1 in 6 in vitro–grown transformants and 4 control plants. Mal d 1 was separated by SDS-PAGE. For detection, radiolabeled goat antibody against mouse IgG or sheep antibody against human IgE was used, respectively. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

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