Cell-free protein synthesis system.

Cell-free protein synthesis system is an extraterrogenous mRNA or DNA-based system that replenishes substrates and energy in the enzyme system of cell extracts to synthesize proteins.
, the in vitro cellless synthesis system has many advantages compared with the traditional in vivo recombination expression system.
For example, special proteins that can express toxic effects on cells or contain non-natural amino acids such as D-amino acids can be used directly as templates to synthesize multiple proteins in parallel, conducting high-volume drug screening and proteomics studies.
cellless protein synthesis system as an open protein expression, its protein expression can reach the milligram level, and due to the short expression cycle, easy to operate, in recent years has been used to express a large number of membrane protein and antibacterial peptides.
Professor Chen Haiqin of jiangnan University's School of Food studied the key membrane binding enzyme- omega-3 fatty acid desaturation enzyme in the synthesis pathway of spore-like fungus alpine spore mold, and constructed the expression vector PIVEXWG1.4-FADS15 suitable for in-body expression of omega-3 fatty acid desaturation enzyme, and realized the efficient expression of the gene by using wheat embryo cellless protein synthesis system.
same time, by adding liposomes to the process of cell-free protein synthesis in wheat embryos, the expressed membrane protein is correctly positioned to the lipid phospholipid bimole molecular layer in order to correctly fold the target protein.
The results show that the expression of the target protein reaches 1.8 mg/mL under the experimental conditions, and the purity of the protein can reach more than 90% after the super-centrifugal purification of the iodized sea alcohol density gradient.
source: advances in biotechnology.