Continuous-exchange cell-free protein-synthesizing system: synthesis of HIV-1 antigen Nef

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Item Type:	Article
Title:	Continuous-exchange cell-free protein-synthesizing system: synthesis of HIV-1 antigen Nef
Creators Name:	Chekulayeva, M.N., Kurnasov, O.V., Shirokov, V.A. and Spirin, A.S.
Abstract:	HIV-1 antigen Nef, Green Fluorescent Protein (GFP), and the fusion protein GFP-Nef (“Green Fluorescent Nef”) were synthesized in bacterial cell-free system with continuous supply of substrates and continuous removal of low-molecular-weight products through a dialysis membrane during incubation, the so-called continuous-exchange cell-free (CECF) system. The identity of synthesized proteins was confirmed by electrophoretic mobility, Western blotting, and GFP fluorescence. The system produced several nanomoles (hundreds of μg) of each protein per ml of the reaction mixture. Construction of GFP-fused proteins is considered as a general strategy for visualization and monitoring of cell-free production of the proteins that have no easily testable functions.
Keywords:	Continuous-Exchange Cell-Free (CECF) System, Combined Transcription/Translation, Green Fluorescent Protein, HIV-1 Antigen Nef
Source:	Biochemical and Biophysical Research Communications
ISSN:	0006-291X
Publisher:	Academic Press
Volume:	280
Number:	3
Page Range:	914-917
Date:	26 January 2001
Official Publication:	https://doi.org/10.1006/bbrc.2000.4188
PubMed:	View item in PubMed

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