Improvement ofescherichia coli cellfree system by utilization of cell extract having additional property
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In theEscherichia coli cell-free system, the modification of cell extract can be achieved by preparation of the strains carrying additional property or those being induced with a certain gene expression prior to harvesting. In this study, we analyzed the cell-free system with S30 extract containing T7 RNA polymerases (S30 extract-T7pol) prepared from E.coli BL21(DE3) strain, which includes T7 RNA polymerase from extrinsic genes by IPTG induction, as a model for the improvement of the cell-free system. The fact that a significant degree of mRNA degradation was observed in the cell-free system with S30 extract-T7pol indicates the increase of ribonuclease activity was an unfavorable influence derived from the cell-extract modification process. We also showed that this influence was settled by the addition of an effective ribonuclease inhibitor, such as copper (II) ion, to the reaction mixture.
Index entriesCell-free system copper ion ribonuclease Escherichia coli protein synthesis
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- 8.Wade, H. E. and Robinson, H. K. (1966),Biochem. J. 101, 467–479.Google Scholar
- 9.Reiner, A. M. (1969),J. Bacteriol. 97, 1431–1436.Google Scholar
- 13.Pratt, J. M. (1984) in,Transcription and Translation: A Practical Approach, Hames, B. D. and Higgins, S. J., eds., IRL, New York, pp. 179–209.Google Scholar