Bean Yellow Dwarf Virus replicons for high-level transgene expression in transgenic plants and cell cultures.
A novel stable transgenic plant expression system was developed using elements of the replication machinery of Bean Yellow Dwarf Virus (BeYDV). The system contains two transgenes: 1) The BeYDV replicon vector with an expression cassette flanked by cis-acting DNA elements of BeYDV, and 2) The viral replication initiator protein (Rep) controlled by an alcohol-inducible promoter. When Rep expression was triggered by treatment with ethanol, it induced release of the BeYDV replicon from stably integrated T-DNA and episomal replication to high copy number. Replicon amplification resulted in substantially increased transgene mRNA levels (up to 80-fold) and translation products (up to 10-fold) after induction of Rep expression by ethanol treatment in tobacco NT1 cells and leaves of whole potato plants. Thus, the BeYDV stable transformant replicon system is a powerful tool for plant-based production of recombinant proteins.
Zhang
Xiuren
X
Department of Plant Biology, Cornell University, Ithaca, NY 14853-1801, USA.
Mason
Hugh
H
D001709:Biotechnology / Q000379:methods*; D002478:Cells, Cultured; D004265:DNA Helicases / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D005784:Gene Amplification; D005822:Genetic Vectors; D010942:Plant Viruses / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012093:Replicon / Q000235:genetics; D011198:Solanum tuberosum / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics*; D019076:Transgenes / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D015534:Trans-Activators; C045264:replication initiator protein; D004265:DNA Helicases