L. sativa-based VLP Production Service

L. sativa-based VLP Production Service

CD BioSciences is a leading global company with advanced equipment and experienced staff dedicated to helping our global customers construct virus-like particles (VLPs) from Lactuca sativa (L. sativa) plants based on the geminivirus replication system to help you develop vaccines, nanomaterials and diagnostic reagents.

Overview of L. sativa

L. sativa, commonly known as lettuce, is an annual plant of the family Asteraceae. N. benthamiana is the most common expression host for the construction of VLPs on plant platforms because of its high biologic yield and the large number of expression vectors suitable for N. benthamiana. However, the leaves of N. benthamiana contain high levels of phenolics and toxic alkaloids that contaminate the resin during purification of VLPs and are difficult to separate from VLPs. Lettuce is a vigorous plant that is easy to cultivate and has a high biological yield. However, unlike tobacco, it produces negligible amounts of phenols and alkaloids, thus greatly reducing costs in the downstream processing of VLPs.

Transient expression in Lettuce. (Kanagaraj AP, et al., 2011)

Overview of BeYDV

Bean yellow dwarf virus (BeYDV), a Mastrevirus of the Geminiviridae, has a single-stranded circular DNA genome that can replicate to very high copy numbers through a loop-rolling mechanism. The BeYDV genome consists of a long intergenic region (LIR), a short intergenic region (SIR) and four open reading frames (ORFs). The four open reading frames (ORFs) encode four proteins: the movement protein, the coat protein, and replication associated proteins Rep and RepA. LIR and SIR are both essential cis-acting elements, while Rep is the only viral protein that is required but can be supplied in trans. The BeYDV-based vector effectively increases the level of protein expression on the plant platform.

Our Services

CD BioSciences offers services covering all experiments related to the construction of VLPs through L. sativa, including the construction of vectors containing the target gene, plant transient expression systems (Agrobacterium) and harvesting of L. sativa plants containing VLPs, as well as VLP-Purification.

Workflow of Our Service

Construction of L. sativa Expression Vectors

It all starts with screening for the target gene or viral code and then using our technology to synthesize genetic instructions that the plant can 'read'. The synthesized target gene is inserted into a bacterium, which then carries the information into L. sativa plant cell.

Plant Transient Expression System

Transient expression mediated by Agrobacterium allows for faster and more efficient production of VLPs by the plant. The plant expression vector is transformed into the rooting Agrobacterium and the Agrobacterium then infiltrates into the L. sativa plant. L. sativa plants are placed in carefully controlled greenhouses to rapidly produce VLPs in large quantities.

Harvest Plants Containing VLPs

Once the plants have finished growing, we harvest the plants and mix the leaves with a solution to extract VLPs. VLPs do not contain genetic material, so they are not infectious. Additionally, plants carry a very low risk of introducing foreign human pathogens.

Purification of VLPs

Our purification services for VLPs constructed from plant platforms include PEG precipitation, ultracentrifugation techniques, and affinity chromatography, to name a few. See the VLP-Purification page for details.

Process of Our Service

We have experienced scientists, technicians and advanced equipment to provide our global customers with a fast, and high-quality service at competitive prices. If you are interested in our services, please contact us for more details, and we will respond promptly.

Reference

1. Kanagaraj, A.P.; et al Expression of dengue-3 premembrane and envelope polyprotein in lettuce chloroplasts. Plant Mol Biol. 2011, 76:323-333.