Synthesis of colicin E1 in a cell-free coupled transcription-translation system

Abstract

Protein was synthesized in a cell-free coupled transcription-translation (S30) system on plasmid Col E1 and phage T2 and Sd DNA templates. Phage T2 DNA and closed plasmid DNA proved to be the most active templates. The latter controlled the synthesis of eight individual proteinsin vitro, including the biologically active protein antibiotic colicin. The titer of colicin synthesized in this cell-free system reached 1024 units/ml, two orders of magnitude higher than the titer of the antibiotic in populations of colicinogenic bacteria.