Based on the comprehensive protein engineering platform established through years of experiences, scientists form Creative Biostructure can provide costum Mempro™ protein production services for Bet v1-like superfamily proteins using cell-free expression system. You can count on us all through your project.
Mempro™ cell-free protein production system is the newly developed system to produce integral membrane proteins, which bypasses the limitations that encountered in cell-based protein production system, such as host cell toxicity, low yield and solubilization and purification using detergents. Bet v1, the major birch pollen allergen, is the prototype of pathogenesis-related (PR)-10 protein family causing respiratory allergy. Notably, Bet v1-like protein from birch pollen is a lipocalin-like protein, which has a TATA binding protein (TBP)-like fold that containing beta-alpha-beta(4)-alpha structural domain. Proteins possessing Bet v1-like fold have been found to be involved in lipid transport, pathogenesis, biosynthesis of secondary metabolites, degradation of aromatic compounds, and antibiotic resistance.
Figure 1. The conserved structure of β-α2-β6-α fold of Bet v1 from Betula pendula. (J. Biomol. Struc. Dynam., 2011)
Creative Biostructure scientists are skillful in high-yield Bet v1-like superfamily protein production based on cell-free protein expression system, we can provide various strategies for Mempro™ cell-free protein production, including:
Among Mempro™ cell-free membrane protein production systems, E. coli S30 extract is widely used for Bet v1-like superfamily protein production. Wheat germ extract can provide eukaryotic translation system for Bet v1-like superfamily protein expression. Baculovirus expression system from Autographa californica nucleopolyhedrovirus (AcNPV) is also a major system for Bet v1-like superfamily protein expression. Rabbit reticulocyte lysate is another useful tool for eukaryotic cell-free protein expression. To explore a suitable source of the extract, many factors, such as the availability of materials, protein origin and complexity, downstream processing, etc., should be considered.
With the Mempro™ cell-free protein production platform, Creative Biostructure are capable of expressing, purifying and crystallizing Bet v1-like proteins to facilitate further structural and functional studies.
One challenge with cell-free protein production is that degradation of the DNA by endogenous nucleases. Creative Biostructure can optimize Mempro™ cell-free protein production for Bet v1-like proteins using plasmids because of they are circular and have no end for the exonucleases to attach.
We provide other various Mempro™ membrane protein production services. Please feel free to contact us for a detailed quote.
References:F. Bernhard, et al. (2013). Cell-free expression - making a mark. Cur. Opin. Struct. Biol., 23: 374-380.F. Roth-Walter, et al. (2014). Bet v1 from birch pollen is a lipocalin-like protein acting as Allergen only when devoid of iron by promoting Th2 lymphocytes. J. Biol. Chem., 289: 17416-17421.J. Chakrabortya and T. K. Duttaa (2011). From lipid transport to oxygenation of aromatic compounds: evolution within the bet v1-like superfamily. J. Biomol. Struc. Dynam., 29(1): 67-78.