원문정보
초록
영어
The cultivation of transformed cells has long been used as a standard route for preparing recombinant proteins. However, the recent explosion in the number of the newly identified ORFs is now demanding a high throughput of protein expression that cannot be readily covered by the
present in vivo expression technology. ell-free protein synthesis is attracting renewed attention as
an alternative technique for overcoming the limited throughput of in vivo expression. With the aim
of developing a high-throughput and highly efficient cell-free protein synthesis system directed by
PCR products, in this study, we describe a systematic approach to enhance the stability of RNA
in cell-free extracts. In another study, we discovered that the productivity of the recombinant proteins from the PCR-amplified templates was enhanced remarkably using an optimized ranslation enhancer sequence. The extra amino acid residues derived from the translation enhancer sequence were effectively removed by utilizing appropriate detergents and peptide cleavage enzyme in the reaction mixture. The method presented here enables the parallel expression of authentic proteins at the elevated levels, and thus it will provide a valuable platform for the large-scale translation of genomic information into protein molecules.