Nucleic Acids Research, 1987, Vol. 15, No. 21 8783-8798
© 1987
Articles |
Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates
Department of Chemistry and Biochemistry, University of Colorado Boulder, CO 80309–0215, USA
Received July 13, 1987. Revised October 1, 1987. Accepted October 1, 1987.
A method is described to synthesize small RNAs of defined length and sequence using T7 RNA polymerase and templates of synthetic DNA which contain the T7 promoter. Partially single stranded templates which are base paired only in the –17 to +1 promoter region are just as active in transcription as linear plasmid DNA. Runoff transcripts initiate at a unique, predictable position, but may have one nucleotide more or less on the 3' terminus. In addition to the full length products, the reactions also yield a large amount of smaller oligoribonucleotides in the range from 2 to 6 nucleotides which appear to be the result of abortive initiation events. Variants in the +1 to +6 region of the promoter are transcribed with reduced efficiency but increase the variety of RNAs which can be made. Transcription reaction conditions have been optimized to allow the synthesis of milligram amounts of virtually any RNA from 12 to 35 nucleotides in length.
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C. E. Bullerwell and M. W. Gray In Vitro Characterization of a tRNA Editing Activity in the Mitochondria of Spizellomyces punctatus, a Chytridiomycete Fungus J. Biol. Chem., January 28, 2005; 280(4): 2463 - 2470. [Abstract] [Full Text] [PDF]
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M. Hass, U. Golnitz, S. Muller, B. Becker-Ziaja, and S. Gunther Replicon System for Lassa Virus J. Virol., December 15, 2004; 78(24): 13793 - 13803. [Abstract] [Full Text] [PDF]
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S. GOPALAKRISHNA, V. GUSTI, S. NAIR, S. SAHAR, and R. K. GAUR Template-dependent incorporation of 8-N3AMP into RNA with bacteriophage T7 RNA polymerase RNA, November 18, 2004; 10(11): 1820 - 1830. [Abstract] [Full Text] [PDF]
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