Synthesis and maturation of green fluorescent protein in a cell-free translation system

Vyacheslav A. Kolb, Eugeny V. Makeyev, William W. Ward, Alexander S. Spirin

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

A cell-free translation system producing mature green fluorescent protein (GFP) can be a useful tool for studying the mechanism and kinetics of GFP chromophore formation, as well as for fast protein engineering. We report here that the mature GFP can be formed in the cell-free translation system from E.coli. The synthesis of GFP in the cell-free system reaches a plateau in 30 to 40 min whereas its maturation is completed in 4 h from the beginning of translation. The delay between the GFP synthesis and the chromophore formation in the cell-free system provides the possibility to isolate and to analyse maturation intermediates for elucidation of the modification pathway.

Original languageEnglish (US)
Pages (from-to)1447-1452
Number of pages6
JournalBiotechnology Letters
Volume18
Issue number12
DOIs
StatePublished - 1996

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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