Abstract
Coinjection of T7 promoter-driven plasmids and T7 RNA polymerase (T7 RNAP) into Xenopus oocytes results in robust protein synthesis, due to simultaneous gene transcription-and-translation (TnT) in the oocyte cytoplasm [Geib, S., Sandoz, G., Carlier, E., V. Cornet, Cheynet-Sauvion, V., De Waard, M., 2001. A novel Xenopus oocyte expression system based on cytoplasmic coinjection of T7-driven plasmids and purified T7-RNA polymerase. Receptors Channels 7, 331-343; Tokmakov, A.A., Matsumoto, E., Shirouzu, M., Yokoyama, S., 2006. Coupled cytoplasmic transcription-and-translation--a method of choice for heterologous gene experession in Xenopus oocytes. J. Biotechnol. 122, 5-15]. In the present study, we demonstrate that the TnT reaction of protein synthesis can be reconstituted in cell-free extracts of Xenopus oocytes and eggs. Similar to the reaction in oocytes, the effective coupling of bacteriophage T7 RNAP-mediated transcription with the eukaryotic translation machinery takes place in the Xenopus oocyte and egg extracts. However, the kinetics of protein and RNA production in the extracts are quite different from those observed in oocytes. Potent RNA synthesis in the extracts starts immediately after the addition of T7 promoter-dri...Continue Reading
References
Nov 1, 1992·Analytical Biochemistry·W KudlickiB Hardesty
Oct 1, 1991·Current Opinion in Biotechnology·J D Richter
Dec 11, 1991·Nucleic Acids Research·G Matthews, A Colman
Aug 1, 1991·Journal of Biochemistry·T Kigawa, S Yokoyama
Jan 1, 1991·Cold Spring Harbor Symposia on Quantitative Biology·A W Murray
Feb 1, 1991·Differentiation; Research in Biological Diversity·J RobertH R Kobel
Nov 25, 1988·Science·A S SpirinY B Alakhov
Feb 15, 1994·Analytical Biochemistry·W KudlickiB Hardesty
Sep 1, 1996·Biotechnology Progress·D M Kim, C Y Choi
Apr 5, 2000·Biochimica Et Biophysica Acta·A J Miller, J J Zhou
Jun 19, 2001·Biotechnology and Bioengineering·D M Kim, J R Swartz
Oct 30, 2001·BioTechniques·G A MartinW Mutter
Jan 16, 2002·The Journal of Biological Chemistry·Sandrine GeibMichel De Waard
Nov 1, 2002·Proceedings of the National Academy of Sciences of the United States of America·Tatsuya SawasakiYaeta Endo
Jan 28, 2003·Current Opinion in Chemical Biology·Shigeyuki Yokoyama
Oct 18, 2003·Biotechnology Advances·Yaeta Endo, Tatsuya Sawasaki
Sep 29, 2004·Trends in Biotechnology·Alexander S Spirin
Mar 1, 2005·Trends in Biotechnology·Federico KatzenWieslaw Kudlicki
Oct 6, 2005·Journal of Biotechnology·Alexander A TokmakovShigeyuki Yokoyama
Oct 20, 2005·Journal of Bioscience and Bioengineering·X JiangT Yamane
Citations
Oct 31, 2008·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Rachel K Tittle, Richard I Hume
Oct 28, 2010·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Alexander A Tokmakov, Yasuo Fukami