Protein synthesis by pure translation systems.
We have developed a partially recombinant, cell-free, protein-synthesis system reconstituted solely from those essential elements of the Escherichia coli translation system, termed protein synthesis using recombinant elements (PURE). It provides higher reaction controllability in comparison to crude cell-free protein-synthesis systems for translation studies and biotechnology applications. The PURE system stands out among translation methods in that it provides not only a simple and unique "reverse" purification method of separating the synthesized protein from reaction mixture, but also that the system can be tailor-made according to individual protein requirements. In this paper, two new approaches to obtaining active proteins are described: the use of molecular chaperones, and modification of the reaction conditions. Several possible applications of the PURE system are also discussed.
Shimizu
Yoshihiro
Y
Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Bldg. FSB-4015-1-5 Kashiwanoha, Kashiwa, Chiba Prefecture 277-8562, Japan.
Kanamori
Takashi
T
Ueda
Takuya
T
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000737:chemistry*; D014176:Protein Biosynthesis; D017510:Protein Folding; D011506:Proteins / Q000138:chemical synthesis / Q000302:isolation & purification
D018832:Molecular Chaperones; D011506:Proteins