Protein synthesis by pure translation systems

Yoshihiro Shimizu, Takashi Kanamori, Takuya Ueda

Research output: Contribution to journalArticle

221 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)299-304
Number of pages6
JournalMethods
Volume36
Issue number3
DOIs
Publication statusPublished - 2005 Jul 1
Externally publishedYes

Fingerprint

Proteins
Molecular Chaperones
Biotechnology
Controllability
Escherichia coli
Purification

Keywords

  • Cell-free protein synthesis
  • Disulfide bond formation
  • Molecular chaperone
  • Protein folding
  • Protein selection
  • Translation
  • Unnatural amino acid

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Protein synthesis by pure translation systems. / Shimizu, Yoshihiro; Kanamori, Takashi; Ueda, Takuya.

In: Methods, Vol. 36, No. 3, 01.07.2005, p. 299-304.

Research output: Contribution to journalArticle

Shimizu, Yoshihiro ; Kanamori, Takashi ; Ueda, Takuya. / Protein synthesis by pure translation systems. In: Methods. 2005 ; Vol. 36, No. 3. pp. 299-304.
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