Plant Toxic Protein Expression and Purification Service
Plant toxic proteins have various potential applications, and the first step in studying their functions is protein expression and purification. The main host used for the production of plant toxic proteins is E. coli. However, heterologous plant toxic proteins can interfere with the survival of the host, leading to growth defects and even death. Therefore, exploring special strategies to enhance the production of toxic proteins is a hot topic of current research.
Figure 1. Potential uses of plant toxic proteins. (Kocyigit, E., et al., 2023)
CD BioSciences helps clients identify and predict the presence of plant toxic genes in molecular cloning experiments, control gene expression, and optimize expression systems to enhance the expression of plant toxic proteins. We also combined multiple methods and genetic tools to ensure reliable results.
Service Content
Plant toxic protein expression host modification service
- Strict control of T7 promoter based on arabinose.
- Introduction of T7 phage lysozyme encoding gene to specifically inhibit T7 polymerase activity.
- Introduction of lac I gene to express lac repressor protein.
- Screening for mutant strains adapted to virulence proteins.
| Hosts adapted for expression of plant toxic proteins | |
| BL21 AI | C41(DE3) |
| BL21 (DE3) pLysS | M15 pREP4 |
| BL21 (DE3) pLysE | OverExpress C43 (DE3) |
| Lemo21(DE3) | BL21 Gold pLysS (DE3) |
Plant toxicity gene expression regulation service
We can rigorously control the expression of toxic proteins below the minimum tolerated concentration of the host or induce the expression of proteins after the host has grown to a certain density. The methods we use include
- Antisense RNA
- CRISPR-dCas9 mediated interference
- Manipulating copy number
Expression service using fusion chaperones
| Fusion chaperone type | Details |
| Solubilizing vectors | Thioredoxin, glutathione transferase (GST), Small ubiquitin-related modifier (SUMO). |
| Aggregation-promoting vectors | PurF fragment, ketone body steroid isomerase, PaP3.30, TAF12 histone folding domain. |
| Self-Lysing Vector | Sce-VMA-intein, Ssp-dnaB-derived mini-intein, fusion tag derived from swine fever virus N-terminal auto-protease Npro. |
Culture conditions optimization service
- Optimization of medium concentration, pH.
- Adding growth factors or cell cycle regulators to the culture medium.
- Screening for appropriate culture temperature.
Plant toxic protein expression and purification services
We transform constructed vectors into suitable hosts for plant toxic protein expression. Then, we provide protein purification services based on
- Affinity Chromatography
- Size exclusion chromatography (SEC)
- Ion exchange chromatography
- Hydrophobic Interaction Chromatography (HIC)
| Types of plant kinases we can express and purify | |
| Ribosome Inactivating Protein (RIP) | Arcelins |
| Lectin | Antimicrobial peptides |
| protease inhibitors and α-amylase inhibitors | Pore-forming toxins |
| Canatoxin-like proteins and ureases | |
CD BioSciences is a biotechnology company focused on plant protein research, providing a variety of related services and products for environmental and energy solutions. If you are interested in our services, please contact us for more details.
References
- Dang, L., & Van Damme, E. J. M. (2015). Toxic proteins in plants. Phytochemistry. 117:51–64.
- Saïda, F., et al. (2006). Expression of highly toxic genes in E. coli: special strategies and genetic tools.Current protein & peptide science. 7(1):47–56.
- Kocyigit, E., et al. (2023) Plant Toxic Proteins: Their Biological Activities, Mechanism of Action and Removal Strategies. Toxins. 15(6):356.
For research use only, not for clinical use.