Dissecting limiting factors of the Protein synthesis Using Recombinant Elements (PURE) system.

ABSTRACT: Reconstituted cell-free protein synthesis systems such as the Protein synthesis Using Recombinant Elements (PURE) system give high-throughput and controlled access to in vitro protein synthesis. Here we show that compared with the commercial S30 crude extract based RTS 100 E. coli HY system, the PURE system has less mRNA degradation and produces up to ∼6-fold full-length proteins. However the majority of polypeptides PURE produces are partially translated or inactive since the signal from firefly luciferase (Fluc) translated in PURE is only ∼2/3^rd of that measured using the RTS 100 E. coli HY S30 system. Both of the 2 batch systems suffer from low ribosome recycling efficiency when translating proteins from 82 k_D to 224 k_D. A systematic

SUBMITTER: Li J 

PROVIDER: S-EPMC5501384 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature