tailieunhanh - Báo cáo khoa học: Ca2+-binding allergens from olive pollen exhibit biochemical and immunological activity when expressed in stable transgenic Arabidopsis

Employing transgenic plants as alternative systems to the conventional Escherichia coli, Pichia pastorisor baculovirus hosts to produce recombin-ant allergens may offer the possibility of having available edible vaccines in the near future. In this study, two EF-hand-type Ca 2+ -binding allergens from olive pollen, Ole e 3 and Ole e 8, were produced in transgenic Arabid-opsis thaliana plants. | ỊFEBS Journal Ca2 -binding allergens from olive pollen exhibit biochemical and immunological activity when expressed in stable transgenic Arabidopsis Amalia Ledesma1 Veronica Moral2 Mayte Villalba1 Julio Salinas2 and Rosalia Rodríguez1 1 Dpto. Bioquimica y Biologia Molecular I Universidad Complutense Madrid Spain 2 Dpto. de Biotecnologia Instituto Nacionalde Investigacion yTecnologia Agraria y Alimentaria Madrid Spain Keywords allergen Ole e 3 Ole e 8 olive pollen plant-expression Correspondence R. Rodríguez Departamento de Bioquimica y Biologia Molecular I Facultad de Ciencias Quimicas Universidad Complutense 28040 Madrid Spain Fax 34 913 944159 Tel. 34 913 944260 E-mail rrg@. Received 29 May 2006 accepted 13 July 2005 doi Employing transgenic plants as alternative systems to the conventional Escherichia coli Pichia pastoris or baculovirus hosts to produce recombinant allergens may offer the possibility of having available edible vaccines in the near future. In this study two EF-hand-type Ca2 -binding allergens from olive pollen Ole e 3 and Ole e 8 were produced in transgenic Arabid-opsis thaliana plants. The corresponding cDNAs under the control of the constitutive CaMV 35S promoter were stably incorporated into the Arabidopsis genome and encoded recombinant proteins AtOle e 3 and AtOle e 8 which exhibited the molecular properties . MS analyses and CD spectra of their olive and or E. coli counterparts. Calcium-binding assays which were carried out to assess the biochemical activity of AtO-le e 3 and AtOle e 8 gave positive results. In addition their mobilities on SDS PAGE were according to the conformational changes derived from their Ca2 -binding capability. The immunological behaviour of Arabidop-sis-expressed proteins was equivalent to that of the natural- and or E. coli-derived allergens as shown by their ability to bind allergen-specific rabbit IgG antiserum and IgE from sensitized patients. These results .

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