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

✍ Scribed by Vyacheslav A. Kolb; Eugeny V. Makeyev; William W. Ward; Alexander S. Spirin

Publisher: Springer Netherlands
Year: 1996
Tongue: English
Weight: 375 KB
Volume: 18
Category: Article
ISSN: 0141-5492
DOI: 10.1007/bf00129353

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✦ Synopsis

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 cellfree 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 cellfree system provides the possibility to isolate and to analyse maturation intermediates for elucidation of the modification pathway.

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