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...
| Authors: | , , , |
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| Format: | article |
| Status: | Published version |
| Publication Date: | 2009 |
| Country: | España |
| Institution: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repository: | DIGITAL.CSIC. Repositorio Institucional del CSIC |
| OAI Identifier: | oai:digital.csic.es:10261/344157 |
| Online Access: | http://hdl.handle.net/10261/344157 |
| Access Level: | Open access |
| Keyword: | Amplicon Controlled expression Expression vector Plum pox virus Potyvirus Virus vector |
| Summary: | 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. |
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