Tobacco Mosaic Virus (TMV) Vector System Service
Creative BioMart Vir-Sci is a worldwide leader in both designing and engineering TMV based expression vectors. By using highly standardized and efficient methods, we permit high quality vectors used for scale up expression of exogenous proteins.
TMV Based Expression System
TMV is one of the most popular plant viruses used for designing expression vector. It has a single-stranded positive-sense RNA (+ssRNA) encapsidated by over 2000 coat protein (CP) molecules. Historically, two main strategies were applied to develop a TMV-based expression vector. First generation vectors are manipulated to express foreign proteins in a “full virus” where the introduced coding sequences are driven by a native viral promoter. For this kind of vector, all necessary elements required for the normal replication cycle are retained. Subsequently, the second generation vector, also designated as the deconstructed vector strategy, was developed to produce recombinant proteins in plants. For this strategy, the genome of TMV was split into several separate modules: one module contains minimum viral elements required for the vector replication, whereas the others contain expression cassettes designed for introduction of exogenous genes. These modules of the recombinant virus vector can be flanked by the left and right T-DNA border sequences from A. tumefaciens, which enables efficient transduction into plants via the Agrobacteria mediated T-DNA transfer mechanism. The magnICON system, composed of three modules, represents one of the most popular deconstructed viral vector. In this scenario, the 5’ module contains the Arabidopsis actin 2 (ACT2) promoter followed by the TMV polymerase and movement protein genes (incorporated with a native CP subgenomic promoter), the 3’ module contains coding sequences of interest and also the terminator, while the third module encodes the PhiC31 integrase acting to recombine the 5’ and 3’ modules. Following the recombination event, one complete TMV-based replicon will be assembled where the exogenous genes are under the control of the subgenomic promoter. This strategy has so far been used for production of a variety of vaccines and other biopharmaceutical proteins.
Features
- TMV-based vectors are compatible with Agrobacteria mediated agroinfiltration, which facilitates efficient infection in a wide range of plants.
- Relatively large size of insertions is tolerated due to deletions of unnecessary native elements.
- Optimization of the TMV genome enables efficient assembly of active replicons.
- Multiple 3’ modules with different targeting sequences can be used simultaneously to direct the expression of foreign genes in different cellular compartments.
- Different subgenomic promoters can be used to express oligomeric proteins from the same vector.
Figure 1. Simplified diagram of deconstructed TMV vectors for recombinase-mediated in planta assembly of viral replicons. RS: recombination site; LB: left border; RB: right border; MP: movement protein.
Service Advantages
- Smart design of deconstructed vector
- Highly optimized protocol
- Top quality vector with flexible production scale
- Strict QC testing
- Competitive price in the market
Workflow of Construction of TMV Based Vectors
The expert team in Creative BioMart Vir-Sci will always find the suitable solution to your research goal. If you are interested in this service, please feel free to contact us for detail information.
References
- Marillonnet, S.; et al. In planta engineering of viral RNA replicons: efficient assembly by recombination of DNA modules delivered by Agrobacterium. Proceedings of the National Academy of Sciences. 2004, 101(18): 6852-6857.
- Cañizares, M. C.; et al. Use of viral vectors for vaccine production in plants. Immunology and Cell Biology. 2005, 83(3): 263-270.