Nucleic Acids Research, 1984, Vol. 12, No. 18 7035-7056
© 1984
MOLECULAR BIOLOGY |
Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter
Department of Biochemistry and Molecular Biology, Harvard University 7 Divinity Avenue, Cambridge, MA 02138, USA
Received June 27, 1984. Revised September 17, 1984. Accepted September 17, 1984.
A simple and efficient method for synthesizing pure single stranded RNAs of virtually any structure is described. This in vitro transcription system is based on the unusually specific RNA synthesis by bacteriophage SP6 RNA polymerase which initiates transcription exclusively at an SP6 promoter. We have constructed convenient cloning vectors that contain an SP6 promoter immediately upstream from a polylinker sequence. Using these SP6 vectors, optimal conditions have been established for in vitro RNA synthesis. The advantages and uses of SP6 derived RNAs as probes for nucleic acid blot and solution hybridizations are demonstrated. We show that single stranded RNA probes of a high specific activity are easy to prepare and can significantly increase the sensitivity of nucleic acid hybridization methods. Furthermore, the SP6 transcription system can be used to prepare RNA substrates for studies on RNA processing (1,5,9) and translation (see accompanying paper).
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H. Kontos, S. Napthine, and I. Brierley Ribosomal Pausing at a Frameshifter RNA Pseudoknot Is Sensitive to Reading Phase but Shows Little Correlation with Frameshift Efficiency Mol. Cell. Biol., December 15, 2001; 21(24): 8657 - 8670. [Abstract] [Full Text] [PDF]
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K. Red-Horse, P. M. Drake, M. D. Gunn, and S. J. Fisher Chemokine Ligand and Receptor Expression in the Pregnant Uterus : Reciprocal Patterns in Complementary Cell Subsets Suggest Functional Roles Am. J. Pathol., December 1, 2001; 159(6): 2199 - 2213. [Abstract] [Full Text] [PDF]
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 M. Burnay, G. Crambert, S. Kharoubi-Hess, K. Geering, and J.-D. Horisberger Bufo marinus bladder H-K-ATPase carries out electroneutral ion transport Am J Physiol Renal Physiol, November 1, 2001; 281(5): F869 - F874. [Abstract] [Full Text] [PDF]
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A. Flores-Morales, N. Stahlberg, P. Tollet-Egnell, J. Lundeberg, R. L. Malek, J. Quackenbush, N. H. Lee, and G. Norstedt Microarray Analysis of the in Vivo Effects of Hypophysectomy and Growth Hormone Treatment on Gene Expression in the Rat Endocrinology, July 1, 2001; 142(7): 3163 - 3176. [Abstract] [Full Text] [PDF]
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 P. M. Drake, M. D. Gunn, I. F. Charo, C.-L. Tsou, Y. Zhou, L. Huang, and S. J. Fisher Human Placental Cytotrophoblasts Attract Monocytes and Cd56bright Natural Killer Cells via the Actions of Monocyte Inflammatory Protein 1{alpha} J. Exp. Med., May 21, 2001; 193(10): 1199 - 1212. [Abstract] [Full Text] [PDF]
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R. W. Oppenheim, S. Wiese, D. Prevette, M. Armanini, S. Wang, L. J. Houenou, B. Holtmann, R. Gotz, D. Pennica, and M. Sendtner Cardiotrophin-1, a Muscle-Derived Cytokine, Is Required for the Survival of Subpopulations of Developing Motoneurons J. Neurosci., February 15, 2001; 21(4): 1283 - 1291. [Abstract] [Full Text] [PDF]
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P. Tollet-Egnell, A. Flores-Morales, N. Ståhlberg, R. L. Malek, N. Lee, and G. Norstedt Gene Expression Profile of the Aging Process in Rat Liver: Normalizing Effects of Growth Hormone Replacement Mol. Endocrinol., February 1, 2001; 15(2): 308 - 318. [Abstract] [Full Text]
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R. K. Stumm, C. Culmsee, M. K.-H. Schafer, J. Krieglstein, and E. Weihe Adaptive Plasticity in Tachykinin and Tachykinin Receptor Expression after Focal Cerebral Ischemia Is Differentially Linked to GABAergic and Glutamatergic Cerebrocortical Circuits and Cerebrovenular Endothelium J. Neurosci., February 1, 2001; 21(3): 798 - 811. [Abstract] [Full Text] [PDF]
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S. Demolombe, D. Franco, P. de Boer, S. Kuperschmidt, D. Roden, Y. Pereon, A. Jarry, A. F. M. Moorman, and D. Escande Differential expression of KvLQT1 and its regulator IsK in mouse epithelia Am J Physiol Cell Physiol, February 1, 2001; 280(2): C359 - C372. [Abstract] [Full Text] [PDF]
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 D. M. Kurtz, L. Tian, B. A. Gower, T. R. Nagy, C. A. Pinkert, and P. A. Wood Transgenic studies of fatty acid oxidation gene expression in nonobese diabetic mice J. Lipid Res., December 1, 2000; 41(12): 2063 - 2070. [Abstract] [Full Text]
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 D. Franco, M. Campione, R. Kelly, P. S. Zammit, M. Buckingham, W. H. Lamers, and A. F. M. Moorman Multiple Transcriptional Domains, With Distinct Left and Right Components, in the Atrial Chambers of the Developing Heart Circ. Res., November 24, 2000; 87(11): 984 - 991. [Abstract] [Full Text] [PDF]
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I. Cuesta, X. Geng, A. Asenjo, and N. Villanueva Structural Phosphoprotein M2-1 of the Human Respiratory Syncytial Virus Is an RNA Binding Protein J. Virol., November 1, 2000; 74(21): 9858 - 9867. [Abstract] [Full Text]
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O. F. Laterza, L. Taylor, S. Unnithan, L. Nguyen, and N. P. Curthoys Mapping and functional analysis of an instability element in phosphoenolpyruvate carboxykinase mRNA Am J Physiol Renal Physiol, November 1, 2000; 279(5): F866 - F873. [Abstract] [Full Text] [PDF]
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F. C. Bender, M. A. Reymond, C. Bron, and A. F. G. Quest Caveolin-1 Levels Are Down-Regulated in Human Colon Tumors, and Ectopic Expression of Caveolin-1 in Colon Carcinoma Cell Lines Reduces Cell Tumorigenicity Cancer Res., October 1, 2000; 60(20): 5870 - 5878. [Abstract] [Full Text]
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 M. Duarte and A. Videira Respiratory Chain Complex I Is Essential for Sexual Development in Neurospora and Binding of Iron Sulfur Clusters Are Required for Enzyme Assembly Genetics, October 1, 2000; 156(2): 607 - 615. [Abstract] [Full Text]
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 J. A. Sullivan and J. C. Gray The Pea light-independent photomorphogenesis1 Mutant Results from Partial Duplication of COP1 Generating an Internal Promoter and Producing Two Distinct Transcripts PLANT CELL, October 1, 2000; 12(10): 1927 - 1938. [Abstract] [Full Text]
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 O. F. LATERZA and N. P. CURTHOYS Effect of Acidosis on the Properties of the Glutaminase mRNA pH-Response Element Binding Protein J. Am. Soc. Nephrol., September 1, 2000; 11(9): 1583 - 1588. [Abstract] [Full Text]
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A. B. M. Nilsson, N. Nitescu, Y. Chen, G. S. Guron, N. Marcussen, G. L. Matejka, and P. Friberg IGF-I treatment attenuates renal abnormalities induced by neonatal ACE inhibition Am J Physiol Regulatory Integrative Comp Physiol, September 1, 2000; 279(3): R1050 - R1060. [Abstract] [Full Text] [PDF]
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B. R. Clarke, F. Esumeh, and I. S. Roberts Cloning, Expression, and Purification of the K5 Capsular Polysaccharide Lyase (KflA) from Coliphage K5A: Evidence for Two Distinct K5 Lyase Enzymes J. Bacteriol., July 1, 2000; 182(13): 3761 - 3766. [Abstract] [Full Text]
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M. L. Draper, J. Wang, N. Valego, W. A. Block Jr., and J. C. Rose Effect of renal denervation on renin gene expression, concentration, and secretion in mature ovine fetus Am J Physiol Regulatory Integrative Comp Physiol, July 1, 2000; 279(1): R263 - R270. [Abstract] [Full Text] [PDF]
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O. F. Laterza and N. P. Curthoys Specificity and functional analysis of the pH-responsive element within renal glutaminase mRNA Am J Physiol Renal Physiol, June 1, 2000; 278(6): F970 - F977. [Abstract] [Full Text] [PDF]
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M. K.-H. Schafer, W. J. Schwaeble, C. Post, P. Salvati, M. Calabresi, R. B. Sim, F. Petry, M. Loos, and E. Weihe Complement C1q Is Dramatically Up-Regulated in Brain Microglia in Response to Transient Global Cerebral Ischemia ,2 J. Immunol., May 15, 2000; 164(10): 5446 - 5452. [Abstract] [Full Text] [PDF]
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P. Béguin, U. Hasler, O. Staub, and K. Geering Endoplasmic Reticulum Quality Control of Oligomeric Membrane Proteins: Topogenic Determinants Involved in the Degradation of the Unassembled Na,K-ATPase alpha Subunit and in Its Stabilization by beta Subunit Assembly Mol. Biol. Cell, May 1, 2000; 11(5): 1657 - 1672. [Abstract] [Full Text]
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 D. Vandier, V. Calvez, L. Massade, A. Gouyette, L. Mickley, T. Fojo, and O. Rixe Transactivation of the Metallothionein Promoter in Cisplatin-Resistant Cancer Cells: a Specific Gene Therapy Strategy J Natl Cancer Inst, April 19, 2000; 92(8): 642 - 647. [Abstract] [Full Text] [PDF]
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M. Laverdiere, J. Beaudoin, and A. Lavigueur Species-specific regulation of alternative splicing in the C-terminal region of the p53 tumor suppressor gene Nucleic Acids Res., March 15, 2000; 28(6): 1489 - 1497. [Abstract] [Full Text] [PDF]
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V. L. Bautch, S. D. Redick, A. Scalia, M. Harmaty, P. Carmeliet, and R. Rapoport Characterization of the vasculogenic block in the absence of vascular endothelial growth factor-A Blood, March 15, 2000; 95(6): 1979 - 1987. [Abstract] [Full Text] [PDF]
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