tailieunhanh - Báo cáo khoa học: Expression analysis of the nucleocytoplasmic lectin ‘Orysata’ from rice in Pichia pastoris

TheOryza sativalectin, abbreviated Orysata, is a mannose-specific, jacalin-related lectin expressed in rice plants after exposure to certain stress condi-tions. Expression of a fusion construct containing the rice lectin sequence linked to enhanced green fluorescent protein in Bright Yellow 2 tobacco cells revealed that Orysata is located in the nucleus and the cytoplasm of the plant cell, indicating that it belongs to the class of nucleocytoplasmic jacalin-related lectins. | IFEBS Journal Expression analysis of the nucleocytoplasmic lectin Orysata from rice in Pichia pastoris l ll . 1 l-s . t I I I 2 I l v 3l ni 4 Bassam Al Atalah Elke Fouquaert Dieter Vanderschaeghe Paul Proost Jan Balzarini David F. Smith5 Pierre Rouge6 Yi Lasanajak5 Nico Callewaert2 and Els J. M. Van Damme1 1 Laboratory of Biochemistry and Glycobiology Department of Molecular Biotechnology Ghent University Belgium 2 Unit for MedicalBiotechnology Department for Molecular BiomedicalResearch Ghent Belgium 3 Laboratory of Molecular Immunology Rega Institute for MedicalResearch Katholieke Universiteit Leuven Belgium 4 Laboratory of Virology and Chemotherapy Rega Institute for MedicalResearch Katholieke Universiteit Leuven Belgium 5 Department of Biochemistry Emory University Schoolof Medicine Atlanta GA USA 6 Signaux et Messages Cellulaires chez les Vegetaux Castanet-Tolosan France Keywords antiviral activity glycan array lectin nucleus Orysata Correspondence E. J. M. Van Damme Laboratory of Biochemistry and Glycobiology Department of Molecular Biotechnology Ghent University Coupure links 653 B-9000 Gent Belgium Fax 32 92646219 Tel 32 92646086 E-mail Received 24 December 2010 revised 5 March 2011 accepted 1 April 2011 doi The Oryza sativa lectin abbreviated Orysata is a mannose-specific jacalin-related lectin expressed in rice plants after exposure to certain stress conditions. Expression of a fusion construct containing the rice lectin sequence linked to enhanced green fluorescent protein in Bright Yellow 2 tobacco cells revealed that Orysata is located in the nucleus and the cytoplasm of the plant cell indicating that it belongs to the class of nucleocytoplasmic jacalin-related lectins. Since the expression level of Orysata in rice tissues is very low the lectin was expressed in the methylotrophic yeast Pichia pasto-ris with the Saccharomyces a-factor sequence to direct the recombinant protein into the .