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Syn5 RNA polymerase synthesizes precise run-off RNA products
The enzyme predominantly used for in vitro run-off RNA synthesis is bacteriophage T7 RNA polymerase. T7 RNA polymerase synthesizes, in addition to run-off products of precise length, transcripts with an additional non-base-paired nucleotide at the 3′-terminus (N + 1 product). This contaminating prod...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950665/ https://www.ncbi.nlm.nih.gov/pubmed/24285303 http://dx.doi.org/10.1093/nar/gkt1193 |
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author | Zhu, Bin Tabor, Stanley Richardson, Charles C. |
author_facet | Zhu, Bin Tabor, Stanley Richardson, Charles C. |
author_sort | Zhu, Bin |
collection | PubMed |
description | The enzyme predominantly used for in vitro run-off RNA synthesis is bacteriophage T7 RNA polymerase. T7 RNA polymerase synthesizes, in addition to run-off products of precise length, transcripts with an additional non-base-paired nucleotide at the 3′-terminus (N + 1 product). This contaminating product is extremely difficult to remove. We recently characterized the single-subunit RNA polymerase from marine cyanophage Syn5 and identified its promoter sequence. This marine enzyme catalyses RNA synthesis over a wider range of temperature and salinity than does T7 RNA polymerase. Its processivity is >30 000 nt without significant intermediate products. The requirement for the initiating nucleotide at the promoter is less stringent for Syn5 RNA polymerase as compared to T7 RNA polymerase. A major difference is the precise run-off transcripts with homogeneous 3′-termini synthesized by Syn5 RNA polymerase. Therefore, the enzyme is advantageous for the production of RNAs that require precise 3′-termini, such as tRNAs and RNA fragments that are used for subsequent assembly. |
format | Online Article Text |
id | pubmed-3950665 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-39506652014-03-12 Syn5 RNA polymerase synthesizes precise run-off RNA products Zhu, Bin Tabor, Stanley Richardson, Charles C. Nucleic Acids Res The enzyme predominantly used for in vitro run-off RNA synthesis is bacteriophage T7 RNA polymerase. T7 RNA polymerase synthesizes, in addition to run-off products of precise length, transcripts with an additional non-base-paired nucleotide at the 3′-terminus (N + 1 product). This contaminating product is extremely difficult to remove. We recently characterized the single-subunit RNA polymerase from marine cyanophage Syn5 and identified its promoter sequence. This marine enzyme catalyses RNA synthesis over a wider range of temperature and salinity than does T7 RNA polymerase. Its processivity is >30 000 nt without significant intermediate products. The requirement for the initiating nucleotide at the promoter is less stringent for Syn5 RNA polymerase as compared to T7 RNA polymerase. A major difference is the precise run-off transcripts with homogeneous 3′-termini synthesized by Syn5 RNA polymerase. Therefore, the enzyme is advantageous for the production of RNAs that require precise 3′-termini, such as tRNAs and RNA fragments that are used for subsequent assembly. Oxford University Press 2014-03 2013-11-26 /pmc/articles/PMC3950665/ /pubmed/24285303 http://dx.doi.org/10.1093/nar/gkt1193 Text en © The Author(s) 2013. Published by Oxford University Press. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Zhu, Bin Tabor, Stanley Richardson, Charles C. Syn5 RNA polymerase synthesizes precise run-off RNA products |
title | Syn5 RNA polymerase synthesizes precise run-off RNA products |
title_full | Syn5 RNA polymerase synthesizes precise run-off RNA products |
title_fullStr | Syn5 RNA polymerase synthesizes precise run-off RNA products |
title_full_unstemmed | Syn5 RNA polymerase synthesizes precise run-off RNA products |
title_short | Syn5 RNA polymerase synthesizes precise run-off RNA products |
title_sort | syn5 rna polymerase synthesizes precise run-off rna products |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950665/ https://www.ncbi.nlm.nih.gov/pubmed/24285303 http://dx.doi.org/10.1093/nar/gkt1193 |
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