- Poster Presentation
- Open Access
Increasing the ease and speed of eukaryotic protein expression: a new cell-free in vitro translation system based on Sf insect cell extracts
Microbial Cell Factoriesvolume 5, Article number: P33 (2006)
For researchers looking for fast access to their protein of interest, cell-free expression systems are an attractive option, as they do not require specialized equipment, are open systems, and offer fast screening of expression constructs for expression efficiency, yield, solubility and other criteria (e.g. requirement of cofactors).
A broad range of eukaryotic proteins require posttranslational modifications such as phosphorylation, glycosylation, or signal peptide cleavage to display full functional activity.
We will describe the most important features of a new system for recombinant eukaryotic protein expression that retains the speed and economy inherent to cell-free methods. Data showing the ability of the system to generate complex posttranslational modifications (PTMs), including highly efficient glycosylation of EPO and ORM1 proteins (Figure 1) and phosphorylation of AKt1 kinase (Figure 2) will be presented. The robustness of the system is demonstrated by the expression of membrane proteins (e.g., OGCP) and the synthesis of several proteins, such as human clotting factors, that could not be expressed in E. coli systems.
Expressed proteins with the EasyXpress Protein Synthesis Insect Kit are post-translationally modified with high efficiency.