חיפוש מתקדם
Journal of Virological Methods
Haviv, S., S. Tolkowsky Laboratory, Department of Virology, Volcani Center, Bet Dagan 50250, Israel
Galiakparov, N., S. Tolkowsky Laboratory, Department of Virology, Volcani Center, Bet Dagan 50250, Israel
Goszczynski, D.E., Virology Unit, Agricultural Research Council, Plant Protection Research Institute, Pretoria, South Africa
Batuman, O., S. Tolkowsky Laboratory, Department of Virology, Volcani Center, Bet Dagan 50250, Israel
Czosnek, H., Robert H. Smith Institute of Plant Science and Genetics in Agriculture, Hebrew University of Jerusalem, Rehovot 76100, Israel
Mawassi, M., S. Tolkowsky Laboratory, Department of Virology, Volcani Center, Bet Dagan 50250, Israel
Grapevine virus A (GVA), a species of the genus Vitivirus, consists of a ∼7.4 kb single-stranded RNA genome of positive polarity, organized into five open reading frames (ORFs). In addition to grape varieties, GVA infects Nicotiana benthamiana plants and protoplasts. We engineered the genome of GVA as a vector that includes duplication of homologous sequences that contain the promoter of the movement protein (MP) sgRNA, supplemented by enzymatic restriction sites to be used as a convenient tool for transient expression of foreign genes from an individual sgRNA. The resulting vector was able to infect and to move in N. benthamiana plants in a manner similar to the wild-type GVA, but it was not stable and the inserted sequence was lost from the genome. Replacing the duplicated promoter with a GVA-MP promoter derived from a distantly related isolate of GVA improved the stability of the inserted sequence. The resulting vector was successfully used to express the reporter gene beta-glucuronidase (GUS) and the coat protein gene of Citrus tristeza virus in inoculated N. benthamiana plants. Development of a useful GVA vector is expected to find a use as a biotechnological tool for improvement of grapevines and it may enable vine breeders to bypass obstacles involved in genetic manipulation of perennial and fruiting plants. © 2005 Elsevier B.V. All rights reserved.
פותח על ידי קלירמאש פתרונות בע"מ -
הספר "אוצר וולקני"
אודות
תנאי שימוש
Engineering the genome of Grapevine virus A into a vector for expression of proteins in herbaceous plants
132
Haviv, S., S. Tolkowsky Laboratory, Department of Virology, Volcani Center, Bet Dagan 50250, Israel
Galiakparov, N., S. Tolkowsky Laboratory, Department of Virology, Volcani Center, Bet Dagan 50250, Israel
Goszczynski, D.E., Virology Unit, Agricultural Research Council, Plant Protection Research Institute, Pretoria, South Africa
Batuman, O., S. Tolkowsky Laboratory, Department of Virology, Volcani Center, Bet Dagan 50250, Israel
Czosnek, H., Robert H. Smith Institute of Plant Science and Genetics in Agriculture, Hebrew University of Jerusalem, Rehovot 76100, Israel
Mawassi, M., S. Tolkowsky Laboratory, Department of Virology, Volcani Center, Bet Dagan 50250, Israel
Engineering the genome of Grapevine virus A into a vector for expression of proteins in herbaceous plants
Grapevine virus A (GVA), a species of the genus Vitivirus, consists of a ∼7.4 kb single-stranded RNA genome of positive polarity, organized into five open reading frames (ORFs). In addition to grape varieties, GVA infects Nicotiana benthamiana plants and protoplasts. We engineered the genome of GVA as a vector that includes duplication of homologous sequences that contain the promoter of the movement protein (MP) sgRNA, supplemented by enzymatic restriction sites to be used as a convenient tool for transient expression of foreign genes from an individual sgRNA. The resulting vector was able to infect and to move in N. benthamiana plants in a manner similar to the wild-type GVA, but it was not stable and the inserted sequence was lost from the genome. Replacing the duplicated promoter with a GVA-MP promoter derived from a distantly related isolate of GVA improved the stability of the inserted sequence. The resulting vector was successfully used to express the reporter gene beta-glucuronidase (GUS) and the coat protein gene of Citrus tristeza virus in inoculated N. benthamiana plants. Development of a useful GVA vector is expected to find a use as a biotechnological tool for improvement of grapevines and it may enable vine breeders to bypass obstacles involved in genetic manipulation of perennial and fruiting plants. © 2005 Elsevier B.V. All rights reserved.
Scientific Publication