Summary
A modified continuous-flow, cell-free (CFCF), coupled transcription and translation system was constructed using an Escherichia coli S30 extract containing endogenous T7 RNA polymerase (S30 extract-T7pol), which was prepared from IPTG induced E. coli BL21(DE3) cells. Though the plasmid pTVCT, which harbored both a bla gene and a chloramphenicol acetyl transferase (CAT) encoding gene under the control of a T7 promoter, was used as a template, only CAT protein was continuously produced by this CFCF-coupled system .
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baranov, V. I., Morozov, I. Y., Ortlepp, S. A. and Spirin, A. S. (1989) Gene 84, 463–466.
Endo, Y., Otsuzuki, S., Ito, K., and Miura, K. (1992) J. Biotech. 25, 221–230.
Kudlicki, W., Kramer, G., and Hardesty, B. (1992) Anal. Biochem. 206, 389–393.
Laemmli, U. K. (1970) Nature 227, 680–685.
Nevin, D. E. and Pratt, J. M. (1991) FEBS lett. 291, 259–263.
Pratt, J. M. (1984) in: Transcription and Translation: A Practical Approach (Hames, B. D., and Higgins, S. J. eds.) pp. 179–209, IRL Press, NY.
Seed, B., and Sheen, J. Y. (1988) Gene 67, 271–277.
Spirin, A. S., Baranov, V. I., Ryabova, L. A., Ovodov, S. Y., and Alakhov, Y. B. (1988) Science 242, 1162–1164.
Spirin, A. S. (1991) in: Frontiers in Bioprocessing II (Todd, P., Sikdar, S. K., and Bier, M. eds.) pp. 31–43, American Chemical Society.
Studier, F. W., and Moffatt, B. A. (1986) J. Mol. Biol. 189, 113.
Wade, H. E. and Robinson, H. K. (1966) Biochem. J. 101, 467–479.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nishimura, N., Kitaoka, Y., Mimura, A. et al. Continuous protein synthesis system with Escherichia coli S30 extract containing endogenous T7 RNA polymerase. Biotechnol Lett 15, 785–790 (1993). https://doi.org/10.1007/BF00180142
Issue Date:
DOI: https://doi.org/10.1007/BF00180142