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

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Presentation transcript:

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 364-369 (February 2005) DOI: 10.1016/j.jaci.2004.10.014 Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

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 2005 115, 364-369DOI: (10.1016/j.jaci.2004.10.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

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 2005 115, 364-369DOI: (10.1016/j.jaci.2004.10.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions

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 2005 115, 364-369DOI: (10.1016/j.jaci.2004.10.014) Copyright © 2005 American Academy of Allergy, Asthma and Immunology Terms and Conditions