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Creative Biolabs can provide stable and high-throughput Mempro™ wheat germ extract-based cell-free protein production services.
Mempro™ cell-free protein production in wheat germ is based on the eukaryotic translation apparatus of wheat seed embryos. Creative Biolabs provides the highest translation efficiency among eukaryotic cell-free membrane protein production using wheat germ extract, and has a very high success rate for the expression of soluble membrane proteins with good quality.
Figure 1. Schematic of wheat germ system-based membrane protein production. (FEBS Lett, 2014)
Conventional wheat germ extract contains RNA N-glycosidase tritin and other inhibitors of translation such as thionin, ribonucleases, deoxyribonucleases and proteases. The high stability of the extract compounds allows the extension of reaction times for even weeks with final protein yields of upto 10 mg/mL at optimal conditions. Most commonly asolectin from soybean (a phospholipid mixture) is used to prepare liposomes for utility in the wheat germ system. After completion of the translation reaction, proteoliposomes containing the expressed membrane protein can be easily purified by centrifugation from the wheat germ translation reactions and directly used in functional analysis of the membrane protein as for example shown for Shaker potassium channels. Besides the Shaker ion channels, the phosphoenolpyruvate/phosphate translocator 1 (AtPPT1), and adenine nucleotide transporter Ant1p. Moreover, detergents can be directly used in the translation reaction to prepare soluble membrane proteins.
Mempro™ membrane protein production in wheat germ system has a wide range of advantages, including：
However, there are some disadvantages of wheat germ system: 1). Wheat germ extracts are relatively difficult to prepare as the embryo is surrounded by the endosperm containing high levels of protein and nucleic acids degrading enzymes as well as translation inhibitors. 2). Some background phosphorylation can be observed in further functional studies. 3). Expression of membrane protein is no glycosylation.
Main applications in MemPro™ wheat germ cell-free membrane protein production, but not limited to:
A. Nozawa, et al. (2010). Production of membrane proteins through the wheat-germ
cell-free technology. Methods Mol. Biol., 607: 213-218.
T. Tsuboi, et al. (2008). Wheat germ cell-free system-based production of malaria proteins for discovery of novel vaccine candidates. Infect. Immun., 76(4): 1702-1708.
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