caa a22 4500 475767454 CHVBK 20180406123610.0 cr unu---uuuuu 170329e20001201xx s 000 0 eng 10.1007/s007050070021 doi (NATIONALLICENCE)springer-10.1007/s007050070021 Development and evaluation of a complementation-dependent gene delivery system based on cucumber mosaic virus [Elektronische Daten] [Y. Zhao, J. Hammond, M. E. Tousignant, R. W. Hammond] Summary.:  To engineer cucumber mosaic virus (CMV-Ix) into a gene vector, genome component RNA 3 of the virus was modified and split into two sub-components, RNA 3A and RNA 3B. In RNA 3A, the open reading frame of the movement protein (MP) was replaced by a reporter gene encoding the green fluorescent protein (GFP), to monitor virus replication and movement. In RNA 3B, the coat protein (CP) gene was eliminated and a multiple cloning site (MCS) was created for foreign gene insertion. Each sub-component alone is defective and relies on its companion sub-component to restore full RNA 3 function. The vector system was evaluated for its ability to deliver and express the bacterial ß-glucuronidase (GUS) gene and a modified bean yellow mosaic virus coat protein (BYMV-CP) gene in Nicotiana benthamiana plants. Results showed that the engineered virus was able to move from cell to cell in the inoculated leaf and enter the minor veins of the inoculated leaf. Foreign gene expression was detected in the inoculated leaves. However, intermolecular recombination between RNA 3A and 3B occurred frequently, preventing efficient systemic expression of the foreign gene(s). Modifications and further evaluations are being undertaken to improve the gene delivery system. Springer-Verlag/Wien, 2000 Zhao Y. Molecular Plant Pathology Laboratory, Plant Sciences Institute, USDA-ARS, Beltsville, Maryland, U.S.A., US aut Hammond J. Floral and Nursery Plants Research Unit, United States National Arboretum, United States Department of Agriculture, Agricultural Research Service, Beltsville, Maryland, U.S.A., US aut Tousignant M. E. Molecular Plant Pathology Laboratory, Plant Sciences Institute, USDA-ARS, Beltsville, Maryland, U.S.A., US aut Hammond R. W. Molecular Plant Pathology Laboratory, Plant Sciences Institute, USDA-ARS, Beltsville, Maryland, U.S.A., US aut Archives of Virology Springer-Verlag 145/11(2000-12-01), 2285-2295 0304-8608 145:11<2285 2000 145 705 https://doi.org/10.1007/s007050070021 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s007050070021 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhao Y. Molecular Plant Pathology Laboratory, Plant Sciences Institute, USDA-ARS, Beltsville, Maryland, U.S.A., US aut NATIONALLICENCE 700 1- Hammond J. Floral and Nursery Plants Research Unit, United States National Arboretum, United States Department of Agriculture, Agricultural Research Service, Beltsville, Maryland, U.S.A., US aut NATIONALLICENCE 700 1- Tousignant M. E. Molecular Plant Pathology Laboratory, Plant Sciences Institute, USDA-ARS, Beltsville, Maryland, U.S.A., US aut NATIONALLICENCE 700 1- Hammond R. W. Molecular Plant Pathology Laboratory, Plant Sciences Institute, USDA-ARS, Beltsville, Maryland, U.S.A., US aut NATIONALLICENCE 773 0- Archives of Virology Springer-Verlag 145/11(2000-12-01), 2285-2295 0304-8608 145:11<2285 2000 145 705 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer