A continuous cell-free protein synthesis system for coupled transcription-translation.

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 (congruent to 0.1 mg) being subsequently eluted through the chamber's membrane and then purified.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Kigawa T,Yokoyama S

doi

10.1093/oxfordjournals.jbchem.a123551

subject

Has Abstract

pub_date

1991-08-01 00:00:00

pages

166-8

issue

2

eissn

0021-924X

issn

1756-2651

journal_volume

110

pub_type

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