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Science 25 November 1988:
Vol. 242. no. 4882, pp. 1162 - 1164
DOI: 10.1126/science.3055301
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Articles

Science, Vol 242, Issue 4882, 1162-1164
Copyright © 1988 by American Association for the Advancement of Science

articles

A continuous cell-free translation system capable of producing polypeptides in high yield

AS Spirin, VI Baranov, LA Ryabova, SY Ovodov, and YB Alakhov

Institute of Protein Research, Academy of Sciences, Moscow Region, USSR.

A cell-free translation system has been constructed that uses a continuous flow of the feeding buffer [including amino acids, adenosine triphosphate (ATP), and guanosine triphosphate (GTP)] through the reaction mixture and a continuous removal of a polypeptide product. Both prokaryotic (Escherichia coli) and eukaryotic (wheat embryos, Triticum sp.) versions of the system have been tested. In both cases the system has proven active for long times, synthesizing polypeptides at a high constant rate for tens of hours. With the use of MS2 phage RNA or brome mosaic virus RNA 4 as templates, 100 copies of viral coat proteins per RNA were synthesized for 20 hours in the prokaryotic or eukaryotic system, respectively. With synthetic calcitonin messenger RNA, 150 to 300 copies of calcitonin polypeptide were produced per messenger RNA in both types of continuous translation systems for 40 hours.


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