earticle

영어

While the expression and screening of genetic libraries for protein engineering is generally carried out in microbial cells, the use of living cells sets intrinsic limitations in numerous dimensions including the throughput of process and choice of assay substrates. These limitations associated with cell-based expression screening can be effectively overcome by expressing enzyme libraries in the cell-free protein synthesis system. Not being subject to the constraints of maintaining cell viability and intact cell membranes, cell-free protein synthesis offers greater flexibility in designing expression and screening procedures. In this study, we will discuss different approaches to express and screen genetic libraries in a cell-free synthesis system derived from Escherichia coli. In particular, the discussion will be focussed on the development of effective methods to assay the expression of individual genes constituting the targeted library.