Novel Vector for Generating RNAs with Defined 3′ Ends and Its Use in Antiviral Strategies
A novel transcription system was constructed that allows trimming of 3′ termini of RNA transcripts in E. coli by endogenous RNase P. Here, the sequence of tRNASer from E. coli fused downstream of the target sequence directs posttranscriptional cleavage 3′ of the target sequence. As a first-target MN...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
1998-01-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/98241st05 |
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| author | Andreas Schwienhorst Björn F. Lindemann |
| author_facet | Andreas Schwienhorst Björn F. Lindemann |
| author_sort | Andreas Schwienhorst |
| collection | DOAJ |
| description | A novel transcription system was constructed that allows trimming of 3′ termini of RNA transcripts in E. coli by endogenous RNase P. Here, the sequence of tRNASer from E. coli fused downstream of the target sequence directs posttranscriptional cleavage 3′ of the target sequence. As a first-target MNV11(+), a self-replicating RNA from the Qβ system was subjected to transcription in vivo. Northern blotting experiments revealed that the primary transcript was indeed successfully processed to an RNA of expected length. The RNA released proved to function as an active template for Qβ replicase. Moreover, E. coli cells producing these short-chain replicator molecules no longer supported multiplication of Qβ phages upon infection. Since the novel transcript-trimming system utilizes the endogenous RNase P activity and does not depend on any particular 3′-terminal RNA sequence of target molecules, it may have wide applications for a number of different targets in prokaryotes. Further applications, including those in eukaryotes, are discussed. |
| format | Article |
| id | doaj-art-1dfd96820d0e4aff8c09111d169fc87f |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 1998-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-1dfd96820d0e4aff8c09111d169fc87f2025-08-20T02:25:51ZengTaylor & Francis GroupBioTechniques0736-62051940-98181998-01-0124111612610.2144/98241st05Novel Vector for Generating RNAs with Defined 3′ Ends and Its Use in Antiviral StrategiesAndreas Schwienhorst0Björn F. Lindemann11Max-Planck-Institute for Biophysical Chemistry, Goettingen, Germany1Max-Planck-Institute for Biophysical Chemistry, Goettingen, GermanyA novel transcription system was constructed that allows trimming of 3′ termini of RNA transcripts in E. coli by endogenous RNase P. Here, the sequence of tRNASer from E. coli fused downstream of the target sequence directs posttranscriptional cleavage 3′ of the target sequence. As a first-target MNV11(+), a self-replicating RNA from the Qβ system was subjected to transcription in vivo. Northern blotting experiments revealed that the primary transcript was indeed successfully processed to an RNA of expected length. The RNA released proved to function as an active template for Qβ replicase. Moreover, E. coli cells producing these short-chain replicator molecules no longer supported multiplication of Qβ phages upon infection. Since the novel transcript-trimming system utilizes the endogenous RNase P activity and does not depend on any particular 3′-terminal RNA sequence of target molecules, it may have wide applications for a number of different targets in prokaryotes. Further applications, including those in eukaryotes, are discussed.https://www.future-science.com/doi/10.2144/98241st05 |
| spellingShingle | Andreas Schwienhorst Björn F. Lindemann Novel Vector for Generating RNAs with Defined 3′ Ends and Its Use in Antiviral Strategies BioTechniques |
| title | Novel Vector for Generating RNAs with Defined 3′ Ends and Its Use in Antiviral Strategies |
| title_full | Novel Vector for Generating RNAs with Defined 3′ Ends and Its Use in Antiviral Strategies |
| title_fullStr | Novel Vector for Generating RNAs with Defined 3′ Ends and Its Use in Antiviral Strategies |
| title_full_unstemmed | Novel Vector for Generating RNAs with Defined 3′ Ends and Its Use in Antiviral Strategies |
| title_short | Novel Vector for Generating RNAs with Defined 3′ Ends and Its Use in Antiviral Strategies |
| title_sort | novel vector for generating rnas with defined 3 ends and its use in antiviral strategies |
| url | https://www.future-science.com/doi/10.2144/98241st05 |
| work_keys_str_mv | AT andreasschwienhorst novelvectorforgeneratingrnaswithdefined3endsanditsuseinantiviralstrategies AT bjornflindemann novelvectorforgeneratingrnaswithdefined3endsanditsuseinantiviralstrategies |