Chemically inducible cucumber mosaic virus protein expression system
The invention relates to a novel chemically inducible plant viral amplicon (CMViva) expression system that permits controllable, high level expression of foreign genes in plant hosts. This system employs agro-infiltration of plants to provide a transient production of a protein of interest, such as a human blood protein. This system provides a major advantage over existing plant expression systems because it allows for consistent expression of foreign or heterologous proteins in plant hosts.
1. A plant expression system for the expression of a heterologous gene in a plant or plant cell culture, the plant expression system comprising a cDNA encoding a CMV1a replicase and a cDNA encoding a CMV2a replicase, wherein the cDNA encoding the CMV1a replicase comprises a CMV RNA 1 cDNA with a 5′ nucleotide sequence deletion consisting of deletion of 57 nucleotides from the 5′ end of the CMV RNA 1 cDNA, wherein said nucleotide sequence deletion prevents replication of CMV1a transcripts when a functional replicase is present, and wherein the cDNA encoding the CMV1a replicase is operably linked to a LEX operator and expression of the CMV1a replicase is estradiol-inducible.
2. The expression system of claim 1 , wherein the heterologous gene codes for a human protein.
3. A plant or plant cell comprising the plant expression system of claim 1 .
4. The expression system of claim 1 , wherein said cDNA encoding said CMV2a replicase is operably linked to a CaMV 35S promoter.
5. The expression system of claim 1 , wherein the heterologous gene is operably linked to a CaMV 35S promoter.
6. The expression system of claim 1 further comprising a cDNA encoding a gene silencing suppressor.