Cytoplasmic inhibition of gene expression and expression of a foreign protein in a monocot plant by a plant viral vector

Abstract

The present invention relates to recombinant viral vectors encoding a transcriptional unit, that encodes a fusion protein, or a foreign protein or a gene of interest to be silenced, which can be expressed in a host. The present invention also relates to the use of these recombinant viral vectors to express a fusion protein, a foreign protein, to silence a gene of interest in a host. The present invention also relates to the use of these recombinant viral vectors to screen a cDNA or genomic library in order to correlate a nucleotide sequence with a phenotypic or biochemical change.

1 - 30 . Cancelled.  
     
     
         31 . A viral genome comprising one or more polynucleotide(s), wherein at least one polynucleotide encodes a first promoter operatively linked to a transcriptional unit, wherein the transcription unit encodes a fusion protein, wherein the fusion protein comprises (1) a first viral protein, (2) a second viral protein, and (3) an autoproteolytic peptide, wherein (3) is fused between (1) and (2), wherein at least one polynucleotide encodes a second promoter operatively linked to the ORF encoding a protein of interest.  
     
     
         32 . A viral genome according to  claim 31  wherein the viral genome is the viral genome of a multipartite RNA plant virus.  
     
     
         33 . A viral genome according to  claim 32  wherein the viral genome is the viral genome of a hordeivirus.  
     
     
         34 . A viral genome according to  claim 33  wherein the hordeivirus is a barley stripe mosaic virus.  
     
     
         35 . A viral genome according to  claim 34  wherein the first viral protein is γb.  
     
     
         36 . A viral genome according to  claim 34  or  35  wherein the second viral protein is βb.  
     
     
         37 . A viral genome according to  claim 31  wherein the autoproteolytic peptide comprises the 2A autoproteolytic peptide of a foot and mouth disease virus.  
     
     
         38 . A viral genome according to  claim 31  wherein the autoproteolytic peptide comprises the amino acid sequence depicted by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 9.  
     
     
         39 . A viral genome according to  claim 31  wherein the protein of interest and the fusion protein are encoded on separate polynucleotides.  
     
     
         40 . A viral genome according to  claim 31  wherein the protein of interest and the fusion protein are capable of expression in a plant cell.  
     
     
         41 . A viral genome according to  claim 40  wherein the plant cell is a monocot plant cell.  
     
     
         42 . A viral genome according to  claim 40  wherein the protein of interest and the fusion protein are capable of systemic expression in a plant.  
     
     
         43 . A viral genome according to  claim 42  wherein the plant is a monocot plant.  
     
     
         44 - 67 . Cancelled.