Expression of foreign genes using a replicating polyprotein producing virus vector
AssigneeThe Texas A&M University System
InventorsJames C. Carrington, Valerian V. Dolja
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Abstract
The present invention provides an expression vector adapted for expressing heterologous proteins in plants susceptible to a polyprotein-producing plant virus. The vector utilizes the unique ability of viral polyprotein proteases to cleave heterologous proteins from viral polyproteins. Also provided is a method for expressing heterologous proteins in plants using these unique expression vectors.
Claims9
A plasmid comprising:
- (a) at least one promoter;
- (b) cDNA, wherein said cDNA comprises sequences that code for a replicatable genome of a polyprotein-producing Tabacco Etch Virus, and;
- (c) at least one unique restriction site flanking a 3' terminus of said cDNA.
A plasmid comprising:
- (a) at least one promoter;
- (b) cDNA, wherein said cDNA comprises sequences that code for a replicatable genome of a polyprotein-producing Tobacco Etch Virus, and wherein said polyprotein comprises at least one protein non-native to the Tobacco Etch Virus, and;
- (c) at least one unique restriction site flanking a 3' terminus of said cDNA.
A method for producing a virus, said method comprising:
- (a) reverse transcribing a polyprotein-producing Tobacco Etch Virus RNA genome into cDNA;
- (b) introducing at least one unique restriction site flanking a 3' terminus of said cDNA;
- (c) inserting said cDNA into a cloning vehicle;
- (d) producing RNA transcripts from said cDNA using an RNA polymerase;
- and
- (e) inoculating plants or plant cells with said RNA transcripts.
A method for producing a virus, said method comprising:
- (a) reverse transcribing a polyprotein-producing tobacco Etch Virus RNA genome into a first cDNA;
- (b) introducing at least one unique restriction site flanking a 3' terminus of said first cDNA;
- (c) inserting into said first cDNA a second cDNA sequence wherein said second cDNA sequence codes for a protein non-native to the Tobacco Etch Virus;
- (d) inserting said first and second cDNA into a cloning vehicle;
- (e) producing RNA transcripts from said cDNA using an RNA polymerase;
- and
- (f) inoculating plants or plant cells with said RNA transcripts.
A method for expressing at least one protein in a plant or plant cell, said method comprising infecting a plant or plant cell susceptible to a polyprotein-producing Tobacco Etch Virus with said Tobacco Etch Virus, expressing said Tobacco Etch Virus to produce said polyprotein, wherein said Tobacco Etch Virus codes for at least one protein non-native to the Tobacco Etch Virus and wherein said non-native protein is released from said polyprotein by proteolytic processing.
Dependencies are inferred from the claim text.