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RNA Template-Directed RNA Synthesis by T7 RNA Polymerase
Christian Cazenave and Olke C. Uhlenbeck
Proceedings of the National Academy of Sciences of the United States of America
Vol. 91, No. 15 (Jul. 19, 1994), pp. 6972-6976
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/2365201
Page Count: 5
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In an attempt to synthesize an oligoribonucleotide by run-off transcription by bacteriophage T7 RNA polymerase, a major transcript was produced that was much longer than expected. Analysis of the reaction indicated that the product resulted from initial DNA-directed run-off transcription followed by RNA template-directed RNA synthesis. This reaction occurred because the RNA made from the DNA template displayed self-complementarity at its 3' end and therefore could form an intra- or intermolecular primed template. In reactions containing only an RNA template, the rate of incorporation of NTPs was quite comparable to DNA-dependent transcription. RNA template-directed RNA synthesis has been found to occur with a great number of oligoribonucleotides, even with primed templates that are only marginally stable. In one instance, we observed a multistep extension reaction converting the oligonucleotide into a final product longer than twice its original length. Presumably, such a process could have generated some of the RNAs found to be efficiently replicated by T7 RNA polymerase.
Proceedings of the National Academy of Sciences of the United States of America © 1994 National Academy of Sciences