A temperature-controlled amplicon system derived from Plum pox potyvirus

The control of replication can facilitate a viral amplicon to reach high expression levels by enabling the virus to escape host defence mechanisms and reducing the deleterious effects of viral infection. We have developed a novel system to regulate amplicon expression by controlling the temperature...

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Detalles Bibliográficos
Autores: Dujovny, Gabriela, Valli, Adrián, Calvo, Maria V., García, Juan Antonio
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2009
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/344157
Acceso en línea:http://hdl.handle.net/10261/344157
Access Level:acceso abierto
Palabra clave:Amplicon
Controlled expression
Expression vector
Plum pox virus
Potyvirus
Virus vector
Descripción
Sumario:The control of replication can facilitate a viral amplicon to reach high expression levels by enabling the virus to escape host defence mechanisms and reducing the deleterious effects of viral infection. We have developed a novel system to regulate amplicon expression by controlling the temperature of plant growth. Nicotiana benthamiana plants were transformed at two different temperatures with a cDNA copy of the Plum pox potyvirus genome harbouring the open reading frame 2 of Porcine circovirus 2 between the nuclear inclusion protein b and coat protein coding sequences. Although transformation at 27 °C mainly yielded nonexpressing amplicons, lines with a tight control of amplicon expression were obtained. Viral replication was not detected in these plants when germinated at 28 °C, but was observed when the plants were shifted to 20 °C. In lines transformed at 24 °C, although the amplicon was expressed at 28 °C, viral accumulation was low and caused only minor growing defects. Viral replication was enhanced in these plants by shifting the temperature to 20 °C; under such conditions, the amplicon reached higher and more persistent expression levels than in plants transformed at 27 °C. These results demonstrate the utility of temperature regulation to control viral amplicon expression.