A Continuous Cell-Free Protein Synthesis System for Coupled Transcription-Translation1
- 1 August 1991
- journal article
- research article
- Published by Oxford University Press (OUP) in The Journal of Biochemistry
- Vol. 110 (2) , 166-168
- https://doi.org/10.1093/oxfordjournals.jbchem.a123551
Abstract
A continuous cell-free protein synthesis system [Spirin et al. (1988) Science 242, 1162–1164] was modified so as to be suitable for coupled transcription-translation, a process useful for obtaining products of cloned genes or cDNAs. A reaction chamber equipped with an ultrafiltration membrane was newly designed and an HPLC pump was used to supply a low molecular weight substrate solution at a constant rate to the viscous reaction mixture in the chamber. By using an Escherichia coli S30 extract in this modified flow system (1 ml), coupled transcription-translation could be continuously performed for 17 h, the synthesized chloramphenicol acetyltransferase ( ≈ 0.1 mg) being subsequently eluted through the chamber's membrane and then purified.Keywords
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