Abstract
Transient expression is a rapid, useful approach for producing proteins of interest in plants. Tobacco mosaic virus (TMV)-based transient expression vectors can express very high levels of foreign proteins in plants. However, TMV vectors are, in general, not efficiently delivered to plant cells by agroinfection. It was determined that agroinfection was very efficient with a 35S promoter-driven TMV replicon that lacked the TMV coat protein gene sequence. This coat protein deletion vector had several useful features as a transient expression system, including improved ease of use, higher protein expression rates, and improved biocontainment. Using this TMV expression vector, some foreign proteins were expressed at levels of 3 to 5 mg/g fresh weight of plant tissue. It is proposed that this new transient expression vector will be a useful tool for expressing recombinant proteins in plants for either research or production purposes.
Highlights
Transient expression is a rapid, useful approach for producing proteins of interest in plants
A recent study has shown that producing recombinant plant proteins in plant cells, instead of yeast (Saccharomyces cerevisiae) or Escherichia coli cells, is more likely to result in the production of properly folded, active plant proteins (Popescu et al, 2007)
We previously constructed an agroinfection-compatible full-length Tobacco mosaic virus (TMV) expression vector called pJL24 (Fig. 1) that expressed all of the TMV genes in addition to a foreign, inserted gene (Lindbo, 2007)
Summary
Transient expression is a rapid, useful approach for producing proteins of interest in plants. If the T-DNA transferred into the plant cell contains a DNA sequence of interest joined to a plant-functional promoter, the transferred DNA will be transcribed in the plant nucleus Because this system is so efficient, easy, and inexpensive to use, it has become a very commonly used strategy for producing proteins in plants (Popescu et al, 2007). Ectopic transient expression of an RNA-silencing suppressor protein (such as the P19 protein from Tomato bushy stunt virus) suppressed the posttranscriptional gene silencing of the cointroduced T-DNA (Voinnet et al, 2003) This resulted in an increase in the amount of recombinant protein expressed. TMV has typically been modified to express foreign genes by either replacing a viral gene (such as the coat protein [CP] gene, for example) with a gene of interest (for review, see Scholthof et al, 1996) or by inserting an additional subgenomic promoter (Dawson et al, 1989; Donson et al, 1991; Pogue et al, 1998) into the viral genome to drive the expression of an inserted foreign gene
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