Nucleic Acids Research, 1984, Vol. 12, No. 18 7057-7070
© 1984
MOLECULAR BIOLOGY |
Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs
Department of Biochemistry and Molecular Biology, Harvard University 7 Divinity Avenue, Cambridge, MA 02138, USA
Received June 27, 1984. Revised September 7, 1984. Accepted September 7, 1984.
We describe a method for the synthesis of microgram quantities of eucaryotic messenger RNAs. Injection into the cytoplasm of frog oocytes and addition to wheat germ extracts show that these synthetic RNAs function efficiently as messenger RNAs. We confirm that a 5' cap on the mRNA is essential for translation in injected oocytes and show that most of the 3' flanking region, including the poly A tail, can be deleted without the abolition of protein synthesis. The method of mRNA synthesis involves in vitro transcription of cDNAs which have been cloned into SP6 vectors (described in the accompanying paper). This method enables one to produce large amounts of mRNA and consequently protein from any cDNA clone.
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X Shi, S Amindari, K Paruchuru, D Skalla, H Burkin, B. Shur, and D. Miller Cell surface beta-1,4-galactosyltransferase-I activates G protein-dependent exocytotic signaling Development, January 3, 2001; 128(5): 645 - 654. [Abstract] [PDF]
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S.-i. Hino, S. Kishida, T. Michiue, A. Fukui, I. Sakamoto, S. Takada, M. Asashima, and A. Kikuchi Inhibition of the Wnt Signaling Pathway by Idax, a Novel Dvl-Binding Protein Mol. Cell. Biol., January 1, 2001; 21(1): 330 - 342. [Abstract] [Full Text]
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R. Dermietzel, M. Kremer, G. Paputsoglu, A. Stang, I. M. Skerrett, D. Gomes, M. Srinivas, U. Janssen-Bienhold, R. Weiler, B. J Nicholson, et al. Molecular and Functional Diversity of Neural Connexins in the Retina J. Neurosci., November 15, 2000; 20(22): 8331 - 8343. [Abstract] [Full Text] [PDF]
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C. Ulens, M. Van Boven, P. Daenens, and J. Tytgat Interaction of p-Fluorofentanyl on Cloned Human Opioid Receptors and Exploration of the Role of Trp-318 and His-319 in {micro}-Opioid Receptor Selectivity J. Pharmacol. Exp. Ther., September 1, 2000; 294(3): 1024 - 1033. [Abstract] [Full Text]
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M. J. Main, J. E. Cryan, J. R. B. Dupere, B. Cox, J. J. Clare, and S. A. Burbidge Modulation of KCNQ2/3 Potassium Channels by the Novel Anticonvulsant Retigabine Mol. Pharmacol., August 1, 2000; 58(2): 253 - 262. [Abstract] [Full Text]
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N. S. Carter, C. B. Mamoun, W. Liu, E. O. Silva, S. M. Landfear, D. E. Goldberg, and B. Ullman Isolation and Functional Characterization of the PfNT1 Nucleoside Transporter Gene from Plasmodium falciparum J. Biol. Chem., March 31, 2000; 275(14): 10683 - 10691. [Abstract] [Full Text] [PDF]
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R. L. Kraus, M. J. Sinnegger, A. Koschak, H. Glossmann, S. Stenirri, P. Carrera, and J. Striessnig Three New Familial Hemiplegic Migraine Mutants Affect P/Q-type Ca2+ Channel Kinetics J. Biol. Chem., March 24, 2000; 275(13): 9239 - 9243. [Abstract] [Full Text] [PDF]
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K. Itoh, A. Antipova, M. J. Ratcliffe, and S. Sokol Interaction of Dishevelled and Xenopus Axin-Related Protein Is Required for Wnt Signal Transduction Mol. Cell. Biol., March 15, 2000; 20(6): 2228 - 2238. [Abstract] [Full Text]
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J. D. Lopinski, J. D. Dinman, and J. A. Bruenn Kinetics of Ribosomal Pausing during Programmed -1 Translational Frameshifting Mol. Cell. Biol., February 15, 2000; 20(4): 1095 - 1103. [Abstract] [Full Text]
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H Weber, C. Symes, M. Walmsley, A. Rodaway, and R. Patient A role for GATA5 in Xenopus endoderm specification Development, January 10, 2000; 127(20): 4345 - 4360. [Abstract] [PDF]
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M Umbhauer, A Penzo-Mendez, L Clavilier, J Boucaut, and J Riou Signaling specificities of fibroblast growth factor receptors in early Xenopus embryo J. Cell Sci., January 8, 2000; 113(16): 2865 - 2875. [Abstract] [PDF]
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A Djiane, J Riou, M Umbhauer, J Boucaut, and D Shi Role of frizzled 7 in the regulation of convergent extension movements during gastrulation in Xenopus laevis Development, January 7, 2000; 127(14): 3091 - 3100. [Abstract] [PDF]
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R Vignali, L Poggi, F Madeddu, and G Barsacchi HNF1(beta) is required for mesoderm induction in the Xenopus embryo Development, January 4, 2000; 127(7): 1455 - 1465. [Abstract] [PDF]
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C.-W. Chiang, N. Carter, W. J. Sullivan Jr., R. G. K. Donald, D. S. Roos, F. N. M. Naguib, M. H. el Kouni, B. Ullman, and C. M. Wilson The Adenosine Transporter of Toxoplasma gondii. IDENTIFICATION BY INSERTIONAL MUTAGENESIS, CLONING, AND RECOMBINANT EXPRESSION J. Biol. Chem., December 3, 1999; 274(49): 35255 - 35261. [Abstract] [Full Text] [PDF]
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J. Li, M. Kogan, S. A. B. Knight, D. Pain, and A. Dancis Yeast Mitochondrial Protein, Nfs1p, Coordinately Regulates Iron-Sulfur Cluster Proteins, Cellular Iron Uptake, and Iron Distribution J. Biol. Chem., November 12, 1999; 274(46): 33025 - 33034. [Abstract] [Full Text] [PDF]
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T. P. Flagg, M. Tate, J. Merot, and P. A. Welling A Mutation Linked with Bartter's Syndrome Locks Kir 1.1a (Romk1) Channels in a Closed State J. Gen. Physiol., November 1, 1999; 114(5): 685 - 700. [Abstract] [Full Text] [PDF]
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J. Wu, L. Zhou, K. Tonissen, R. Tee, and K. Artzt The Quaking I-5 Protein (QKI-5) Has a Novel Nuclear Localization Signal and Shuttles between the Nucleus and the Cytoplasm J. Biol. Chem., October 8, 1999; 274(41): 29202 - 29210. [Abstract] [Full Text] [PDF]
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A. M. Barbieri, G. Lupo, A. Bulfone, M. Andreazzoli, M. Mariani, F. Fougerousse, G. G. Consalez, G. Borsani, J. S. Beckmann, G. Barsacchi, et al. A homeobox gene, vax2, controls the patterning of the eye dorsoventral axis PNAS, September 14, 1999; 96(19): 10729 - 10734. [Abstract] [Full Text] [PDF]
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K. Itoh and S. Y. Sokol Axis determination by inhibition of Wnt signaling in Xenopus Genes & Dev., September 1, 1999; 13(17): 2328 - 2336. [Abstract] [Full Text]
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N. Masuyama, H. Hanafusa, M. Kusakabe, H. Shibuya, and E. Nishida Identification of Two Smad4 Proteins in Xenopus. THEIR COMMON AND DISTINCT PROPERTIES J. Biol. Chem., April 23, 1999; 274(17): 12163 - 12170. [Abstract] [Full Text] [PDF]
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W. J. Hansen, N. J. Cowan, and W. J. Welch Prefoldin-Nascent Chain Complexes in the Folding of Cytoskeletal Proteins J. Cell Biol., April 19, 1999; 145(2): 265 - 277. [Abstract] [Full Text] [PDF]
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B. Saul, T. Kuner, D. Sobetzko, W. Brune, F. Hanefeld, H.-M. Meinck, and C.-M. Becker Novel GLRA1 Missense Mutation (P250T) in Dominant Hyperekplexia Defines an Intracellular Determinant of Glycine Receptor Channel Gating J. Neurosci., February 1, 1999; 19(3): 869 - 877. [Abstract] [Full Text] [PDF]
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S. M. Logan, F. E. Rivera, and J. P. Leonard Protein Kinase C Modulation of Recombinant NMDA Receptor Currents: Roles for the C-Terminal C1 Exon and Calcium Ions J. Neurosci., February 1, 1999; 19(3): 974 - 986. [Abstract] [Full Text] [PDF]
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K Wunnenberg-Stapleton, I. Blitz, C Hashimoto, and K. Cho Involvement of the small GTPases XRhoA and XRnd1 in cell adhesion and head formation in early Xenopus development Development, January 12, 1999; 126(23): 5339 - 5351. [Abstract] [PDF]
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N Ohan, Y Agazie, C Cummings, R Booth, M Bayaa, and X. Liu RHO-associated protein kinase alpha potentiates insulin-induced MAP kinase activation in Xenopus oocytes J. Cell Sci., January 7, 1999; 112(13): 2177 - 2184. [Abstract] [PDF]
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M Hild, A Dick, G. Rauch, A Meier, T Bouwmeester, P Haffter, and M Hammerschmidt The smad5 mutation somitabun blocks Bmp2b signaling during early dorsoventral patterning of the zebrafish embryo Development, January 5, 1999; 126(10): 2149 - 2159. [Abstract] [PDF]
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